Inhibitors of histone deacetylase and prodrugs thereof

ABSTRACT

The invention relates to the inhibition of histone deacetylase. The invention provides compounds, prodrugs thereof, and methods for inhibiting histone deacetylase enzymatic activity. The invention also provides compositions and methods for treating cell proliferative diseases and conditions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority of U.S. Application No.60/870,768, filed Dec. 19, 2006.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to the inhibition of histone deacetylase. Moreparticularly, the invention relates to compounds and prodrugs thereofthat inhibit, histone deacetylase enzymatic activity. The invention alsorelates to methods of inhibiting histone deacetylase enzymatic activity.

2. Summary of the Related Art

In eukaryotic cells, nuclear DNA associates with histones to form acompact complex called chromatin. The histones constitute a family ofbasic proteins which are generally highly conserved across eukaryoticspecies. The core histones, termed H2A, H₂B, H3, and H4, associate toform a protein core. DNA winds around this protein core, with the basicamino acids of the histones interacting with the negatively chargedphosphate groups of the DNA. Approximately 146 base pairs of DNA wraparound a histone core to make up a nucleosome particle, the repeatingstructural motif of chromatin.

Csordas, Biochem. J., 286: 23-38 (1990) teaches that histones aresubject to posttranslational acetylation of the e-amino groups ofN-terminal lysine residues, a reaction that is catalyzed by histoneacetyl transferase (HAT1). Acetylation neutralizes the positive chargeof the lysine side chain, and is thought to impact chromatin structure.Indeed, Taunton et al., Science, 272: 408-411 (1996), teaches thataccess of transcription factors to chromatin templates is enhanced byhistone hyperacetylation. Taunton et al. further teaches that anenrichment in underacetylated histone H4 has been found intranscriptionally silent regions of the genome.

Histone acetylation is a reversible modification, with deacetylationbeing catalyzed by a family of enzymes termed histone deacetylases(HDACs). Grozinger et al., Proc. Natl. Acad. Sci. USA, 96: 4868-4873(1999), teaches that HDACs may be divided into two classes, the firstrepresented by yeast Rpd3-like proteins, and the second represented byyeast Hda1-like proteins.

Histone deacetylases play an important role in gene regulation inmammalian cells. Gray and Ekstrom, Expr. Cell. Res. 262: 75-83 (2001);Zhou et al., Proc. Natl. Acad. Sci. USA 98: 10572-10577 (2001); Kao etal. J. Biol. Chem. 277: 187-193 (2002) and Gao et al. J. Biol. Chem.277: 25748-25755 (2002) teach that there are 11 members of the histonedeacetylase (HDAC) family. Another family of deacetylases involved ingene expression is the Sir2 family. Gray and Ekstrom, supra, teach thatthere are seven members of the Sir2 family in humans.

Class I human histone deacetylases include HDAC1, HDAC2, HDAC3 andHDAC8. The Class I enzymes are expressed in a wide variety of tissuesand are reported to be localized in the nucleus. Class II human histonedeacetylases include HDAC4, HDAC5, HDAC6, HDAC7, HDAC9 and HDAC10. TheClass II enzymes have been described as limited in tissue distributionand they can shuttle between the nucleus and the cytoplasm. The Class IIenzymes are further divided into Class IIa (HDAC4, HDAC5, HDAC7 andHDAC9) and Class IIb (HDAC6 and HDAC10). Recent classifications placeHDAC11 in a class of its own.

Richon et al., Proc. Natl. Acad. Sci. USA, 95: 3003-3007 (1998),discloses that HDAC activity is inhibited by trichostatin A (TSA), anatural product isolated from Streptomyces hygroscopicus, and by asynthetic compound, suberoylanilide hydroxamic acid (SAHA). Yoshida andBeppu, Exper. Cell Res., 177: 122-131 (1988), teaches that TSA causesarrest of rat fibroblasts at the G₁ and G₂ phases of the cell cycle,implicating HDAC in cell cycle regulation. Indeed, Finnin et al.,Nature, 401: 188-193 (1999), teaches that TSA and SAHA inhibit cellgrowth, induce terminal differentiation, and prevent the formation oftumors in mice.

U.S. Pat. No. RE39850, incorporated herein by reference, disclosescompounds that inhibit HDAC activity for intervening in cell cycleregulation and therapeutic potential in the treatment of cellproliferative diseases or conditions. However, these compounds canexhibit poor bioavailability, thereby limiting their therapeuticpotential. It is therefore desirable to also prepare bioavailableanalogs of such active compounds.

SUMMARY OF THE INVENTION

The invention provides compounds, prodrugs and methods for treatingdiseases or conditions ameliorated by modulating HDAC activity, such ascell proliferative diseases, or fungal infection, by administering suchprodrugs. In particular, the invention provides prodrug inhibitors ofhistone deacetylase enzymatic activity. These prodrugs are cleavable(e.g., hydrolysable) in a mammalian cell, a plant cell or fungalpathogen cell. Thus, also included within the scope of the presentinvention are products of the cleaved prodrugs, which include novelhydroxamate based compounds. For the purpose of clarity, a “prodrugcompound” or “prodrug” of the present invention is intended to mean anon-cleaved compound as defined by Formula (1), (2) and (3). A “cleavageproduct” of the prodrug is intended to mean a prodrug compound fromwhich the prodrug moiety has been removed.

In a first aspect, therefore, the invention provides prodrugs ofinhibitors of histone deacetylase, the prodrugs having the formula (1):

Cy-L¹-Ar-Y¹—C(O)—N(R^(x))—Z  (1)

and pharmaceutically acceptable salts thereof, wherein

-   Cy is —H, cycloalkyl, aryl, heteroaryl, or heterocyclyl, any of    which may be optionally substituted;-   L¹ is —(CH₂)_(m)—W—, where m is 0, 1, 2, 3, or 4, and W is selected    from the group consisting of —C(O)NH—, —S(O)₂NH—, —NHC(O)—,    —NHS(O)₂—, and —NH—C(O)—NH—;-   Ar is arylene, wherein said arylene optionally may be additionally    substituted and optionally may be fused to an aryl or heteroaryl    ring, or to a saturated or partially unsaturated cycloalkyl or    heterocyclic ring, any of which may be optionally substituted;-   Y¹ is a chemical bond or a straight- or branched-chain saturated    alkylene, wherein said alkylene may be optionally substituted;-   Z is —R²⁰, —O—R²⁰, —R²¹, or

wherein —R²⁰ is selected from the group consisting of —C(O)—R¹⁰,—C(O)O—R¹⁰, —R¹¹, —CH(R¹²)—O—C(O)—R¹⁰,—C(O)—C[(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³), —S(O₂)R¹⁰, —P(O)(OR¹⁰)(OR¹⁰),—C(O)—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰, —C(O)—O—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰ and—C(O)—(CH₂)_(n)—C(O)OR¹⁰, —C(O)—(CH₂)₁₋₄—C(OH)(COOR¹⁰)—(CH₂)₁₋₄—COOR¹⁰,—C(O)-[C(R¹⁴)(R¹⁴)]₁₋₄—P(O)(OH)(OH),—C(O)—(CH₂)₁₋₄—N(R¹⁴)—C[═N(R^(10′))]-N(R^(10′))(R^(10′)),—C(O)—(CH₂)—CH(OH)—(CH₂)—N(CH₃)(CH₃), —C(O)—CH(NH₂)—(CH₂)₁₋₆—COOH(preferably —C(O)—CH(NH₂)—(CH₂)—COOH), provided that the N to which Z isbound is not directly bonded to two O atoms; and further provided that(a) when Z is —R²⁰ then R^(x) is —OH, and (b) when Z is —OR²⁰ then R^(x)is —H;

-   R^(x) is H or —OH;-   or-   R^(x) is absent and R²⁰ forms an optionally substituted heterocyclic    ring with the N to which it is attached;-   n is 0, 1, 2, 3, or 4, preferably 1, 2, 3, or 4;-   each R¹⁰ is independently selected from the group consisting of    hydrogen, optionally substituted C₁-C₂₀ alkyl, optionally    substituted C₂-C₂₀ alkenyl, optionally substituted C₂-C₂₀ alkynyl,    optionally substituted C₁-C₂₀ alkoxycarbonyl, optionally substituted    cycloalkyl, optionally substituted heterocycloalkyl, optionally    substituted aryl, optionally substituted heteroaryl, optionally    substituted cycloalkylalkyl, optionally substituted    heterocycloalkylalkyl, optionally substituted arylalkyl, optionally    substituted heteroarylalkyl, optionally substituted    cycloalkylalkenyl, optionally substituted heterocycloalkylalkenyl,    optionally substituted arylalkenyl, optionally substituted    heteroarylalkenyl, optionally substituted cycloalkylalkynyl,    optionally substituted heterocycloalkylalkynyl, optionally    substituted arylalkynyl, optionally substituted heteroarylalkynyl,    optionally substituted alkylcycloalkyl, optionally substituted    alkylheterocycloalkyl, optionally substituted alkylaryl, optionally    substituted alkylheteroaryl, optionally substituted    alkenylcycloalkyl, optionally substituted alkenylheterocycloalkyl,    optionally substituted alkenylaryl, optionally substituted    alkenylheteroaryl, optionally substituted alkynylcycloalkyl,    optionally substituted alkynylheterocycloalkyl, optionally    substituted alkynylary, optionally substituted alkynylheteroaryl, a    sugar residue, and an amino acid residue (preferably bonded through    the carboxy terminus of the amino acid);-   each R^(10′) is independently hydrogen or C₁₋₆alkyl, or-   R¹⁰ and R^(10′) together with the carbon atom to which they are    attached form an optionally substituted spirocycloalkyl;-   R²¹ is a sugar or -amino acid-R¹³, wherein R¹³ is covalently bound    to the N-terminus;-   R¹¹ is selected from the group consisting of hydrogen, optionally    substituted heterocycloalkyl, optionally substituted aryl, and    optionally substituted heteroaryl;-   R¹² is selected from hydrogen or alkyl; and-   R¹³ is selected from the group consisting of hydrogen,    —C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl,    —C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-N(R^(10′))(R^(10′))C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-aryl,    —C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-heteroaryl, —C(O)-aryl,    —C(O)-heteroaryl, an amino protecting group, and R¹⁰; and-   each R¹⁴ is independently selected from the group consisting of H,    C₁-C₆alkyl and cycloalkyl, or two R¹⁴, together with the atom to    which they are attached, form a cycloalkyl.

In a second embodiment, the invention provides prodrugs of inhibitors ofhistone deacetylase, the prodrugs having the formula (2):

Cy-L²-Ar-Y²—C(O)N(R^(x))—Z  (2)

and pharmaceutically acceptable salts thereof, wherein

-   Cy is H or is cycloalkyl, aryl, heteroaryl, or heterocyclyl, any of    which may be optionally substituted, provided that Cy is not a    (spirocycloalkyl)heterocyclyl;-   L² is C₁-C₆ saturated alkylene, C₂-C₆ alkenylene or C₂-C₆    alkynylene, wherein the alkylene or alkenylene optionally may be    substituted, and wherein one or two of the carbon atoms of the    alkylene is optionally replaced by a heteroatomic moiety    independently selected from the group consisting of O; NR′, R′ being    alkyl, acyl, or hydrogen; S; S(O); or S(O)₂;-   Ar is arylene, wherein said arylene optionally may be additionally    substituted and optionally may be fused to an aryl or heteroaryl    ring, or to a saturated or partially unsaturated cycloalkyl or    heterocyclic ring, any of which may be optionally substituted;-   Y² is a chemical bond or a straight- or branched-chain saturated    alkylene, which may be optionally substituted, provided that the    alkylene is not substituted with a substituent of the formula —C(O)R    wherein R comprises an α-amino acyl moiety;-   R^(x) is H or —OH;-   Z is —R²⁰, —O—R²⁰, —R²¹, or

wherein —R²⁰ is selected from the group consisting of —C(O)—R¹⁰,—C(O)O—R¹⁰, —R¹¹, —CH(R¹²)—O—C(O)—R¹⁰,—C(O)—C[(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³), —S(O₂)R¹⁰, —P(O)(OR¹⁰)(OR¹⁰),—C(O)—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰,—C(O)—(CH₂)₁₋₄—C(OH)(COOR¹⁰)—(CH₂)₁₋₄—COOR¹⁰,—C(O)-[C(R¹⁴)(R¹⁴)]₁₋₄—P(O)(OH)(OH),—C(O)—(CH₂)₁₋₄—N(R¹⁴)—C[═N(R^(10′))]-N(R^(10′))(R^(10′)),—C(O)—(CH₂)—CH(OH)—(CH₂)—N(CH₃)(CH₃), —C(O)—CH(NH₂)—(CH₂)₁₋₆—COOH(preferably —C(O)—CH(NH₂)—(CH₂)—COOH),—C(O)—O—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰ and —C(O)—(CH₂)_(n)—C(O)OR¹⁰,provided that the N to which Z is bound is not directly bonded to two Oatoms; and further provided that (a) when Z is —R²⁰ then R^(x) is —OH,and (b) when Z is —OR²⁰ then R^(x) is —H;

-   or-   R^(x) is absent and R²⁰ forms an optionally substituted heterocyclic    ring with the N to which it is attached;-   n is 0, 1, 2, 3, or 4, preferably 1, 2, 3, or 4;-   each R¹⁰ is independently selected from the group consisting of    hydrogen, optionally substituted C₁-C₂₀ alkyl, optionally    substituted C₂-C₂₀ alkenyl, optionally substituted C₂-C₂₀ alkynyl,    optionally substituted C₁-C₂₀ alkoxycarbonyl, optionally substituted    cycloalkyl, optionally substituted heterocycloalkyl, optionally    substituted aryl, optionally substituted heteroaryl, optionally    substituted cycloalkylalkyl, optionally substituted    heterocycloalkylalkyl, optionally substituted arylalkyl, optionally    substituted heteroarylalkyl, optionally substituted    cycloalkylalkenyl, optionally substituted heterocycloalkylalkenyl,    optionally substituted arylalkenyl, optionally substituted    heteroarylalkenyl, optionally substituted cycloalkylalkynyl,    optionally substituted heterocycloalkylalkynyl, optionally    substituted arylalkynyl, optionally substituted heteroarylalkynyl,    optionally substituted alkylcycloalkyl, optionally substituted    alkylheterocycloalkyl, optionally substituted alkylaryl, optionally    substituted alkylheteroaryl, optionally substituted    alkenylcycloalkyl, optionally substituted alkenylheterocycloalkyl,    optionally substituted alkenylaryl, optionally substituted    alkenylheteroaryl, optionally substituted alkynylcycloalkyl,    optionally substituted alkynylheterocycloalkyl, optionally    substituted alkynylary, optionally substituted alkynylheteroaryl, a    sugar residue, and an amino acid residue (preferably bonded through    the carboxy terminus of the amino acid);-   each R^(10′) is independently hydrogen or C₁₋₆alkyl, or-   R¹⁰ and R^(10′) together with the carbon atom to which they are    attached form an optionally substituted spirocycloalkyl;-   R²¹ is a sugar or -amino acid-R¹³, wherein R¹³ is covalently bound    to the N-terminus;-   R¹¹ is selected from the group consisting of hydrogen, optionally    substituted heterocycloalkyl, optionally substituted aryl, and    optionally substituted heteroaryl;-   R¹² is selected from hydrogen or alkyl; and-   R¹³ is selected from the group consisting of hydrogen,    —C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl,    —C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-N(R^(10′))(R^(10′)),    —C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-aryl,    —C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-heteroaryl, —C(O)-aryl,    —C(O)-heteroaryl, an amino protecting group, and R¹⁰; and-   each R¹⁴ is independently selected from the group consisting of H,    C₁-C₆alkyl and cycloalkyl, or two R¹⁴, together with the atom to    which they are attached, form a cycloalkyl.

In a third embodiment, the invention provides prodrugs of inhibitors ofhistone deacetylase, the prodrugs having the formula (3):

Cy-L³-Ar-Y³—C(O)N(R^(x))—Z  (3)

and pharmaceutically acceptable salts thereof, wherein

-   Cy is —H, cycloalkyl, aryl, heteroaryl, or heterocyclyl, any of    which may be optionally substituted, provided that Cy is not a    (spirocycloalkyl)heterocyclyl;-   L³ is selected from the group consisting of    -   (a) —(CH₂)_(m)—W—, where m is 0, 1, 2, 3, or 4, and W is        selected from the group consisting of —C(O)NH—, —S(O)₂NH—,        —NHC(O)—, —NHS(O)₂—, and —NH—C(O)—NH—; and    -   (b) C₁-C₆ alkylene or C₂-C₆ alkenylene, wherein the alkylene or        alkenylene optionally may be substituted, and wherein one of the        carbon atoms of the alkylene optionally may be replaced by O;        NR′, R′ being alkyl, acyl, or hydrogen; S; S(O); or S(O)₂;-   Ar is arylene, wherein said arylene optionally may be additionally    substituted and optionally may be fused to an aryl or heteroaryl    ring, or to a saturated or partially unsaturated cycloalkyl or    heterocyclic ring, any of which may be optionally substituted; and-   Y³ is C₂ alkenylene or C₂ alkynylene, wherein one or both carbon    atoms of the alkenylene optionally may be substituted with alkyl,    aryl, alkaryl, or aralkyl;-   R^(x) is H or —OH;-   Z is —R²⁰, —O—R²⁰, —R²¹, or

wherein —R²⁰ is selected from the group consisting of —C(O)—R¹⁰,—C(O)O—R¹⁰, —R¹¹, —CH(R¹²)—O—C(O)—R¹⁰,—C(O)—C[(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³),—S(O₂)R¹⁰—C(O)—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰,—C(O)—(CH₂)₁₋₄—C(OH)(COOR¹⁰)—(CH₂)₁₋₄—COOR¹⁰,—C(O)-[C(R¹⁴)(R¹⁴)]₁₋₄—P(O)(OH)(OH),—C(O)—(CH₂)₁₋₄—N(R¹⁴)—C[═N(R^(10′))]-N(R^(10′))(R^(10′)),—C(O)—(CH₂)—CH(OH)—(CH₂)—N(CH₃)(CH₃), —C(O)—CH(NH₂)—(CH₂)₁₋₆—COOH(preferably —C(O)—CH(NH₂)—(CH₂)—COOH),—C(O)—O—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰ and —C(O)—(CH₂)_(n)—C(O)OR¹⁰,provided that the N to which Z is bound is not directly bonded to two Oatoms; and further provided that (a) when Z is —R²⁰ then R^(x) is —OH,and (b) when Z is —OR²⁰ then R^(x) is —H;

-   or-   R^(x) is absent and R²⁰ forms an optionally substituted heterocyclic    ring with the N to which it is attached;-   n is 0, 1, 2, or 4, preferably 1, 2, 3, or 4;-   each R¹⁰ is independently selected from the group consisting of    hydrogen, optionally substituted C₁-C₂₀ alkyl, optionally    substituted C₂-C₂₀ alkenyl, optionally substituted C₂-C₂₀ alkynyl,    optionally substituted C₁-C₂₀ alkoxycarbonyl, optionally substituted    cycloalkyl, optionally substituted heterocycloalkyl, optionally    substituted aryl, optionally substituted heteroaryl, optionally    substituted cycloalkylalkyl, optionally substituted    heterocycloalkylalkyl, optionally substituted arylalkyl, optionally    substituted heteroarylalkyl, optionally substituted    cycloalkylalkenyl, optionally substituted heterocycloalkylalkenyl,    optionally substituted arylalkenyl, optionally substituted    heteroarylalkenyl, optionally substituted cycloalkylalkynyl,    optionally substituted heterocycloalkylalkynyl, optionally    substituted arylalkynyl, optionally substituted heteroarylalkynyl,    optionally substituted alkylcycloalkyl, optionally substituted    alkylheterocycloalkyl, optionally substituted alkylaryl, optionally    substituted alkylheteroaryl, optionally substituted    alkenylcycloalkyl, optionally substituted alkenylheterocycloalkyl,    optionally substituted alkenylaryl, optionally substituted    alkenylheteroaryl, optionally substituted alkynylcycloalkyl,    optionally substituted alkynylheterocycloalkyl, optionally    substituted alkynylary, optionally substituted alkynylheteroaryl, a    sugar residue, and an amino acid residue (preferably bonded through    the carboxy terminus of the amino acid);-   each R^(10′) is independently hydrogen or C₁₋₆alkyl, or-   R¹⁰ and R^(10′) together with the carbon atom to which they are    attached form an optionally substituted spirocycloalkyl;-   R²¹ is a sugar or -amino acid-R¹³, wherein R¹³ is covalently bound    to the N-terminus;-   R¹¹ is selected from the group consisting of hydrogen, optionally    substituted heterocycloalkyl, optionally substituted aryl, and    optionally substituted heteroaryl;-   R¹² is selected from hydrogen or alkyl; and-   R¹³ is selected from the group consisting of hydrogen,    —C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl,    —C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-N(R^(10′))(R^(10′))C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-aryl,    —C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-heteroaryl, —C(O)-aryl,    —C(O)-heteroaryl, an amino protecting group, and R¹⁰; and-   each R¹⁴ is independently selected from the group consisting of H,    C₁-C₆alkyl and cycloalkyl, or two R¹⁴, together with the atom to    which they are attached, form a cycloalkyl.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The invention provides prodrugs, cleavage products thereof, and methodsfor inhibiting histone deacetylase enzymatic activity. The inventionalso provides compositions and methods for treating diseases orconditions ameliorated by modulating HDAC activity, such as cellproliferative diseases and conditions, and fungal infection. The patentand scientific literature referred to herein establishes knowledge thatis available to those with skill in the art. The issued patents,applications, and references that are cited herein are herebyincorporated by reference to the same extent as if each was specificallyand individually indicated to be incorporated by reference. In the caseof inconsistencies, the present disclosure will prevail.

For purposes of the present invention, the following definitions will beused:

Unless otherwise indicated by context, the terms “histone deacetylase”and “HDAC” are intended to refer to any one of a family of enzymes thatremove acetyl groups from the ε-amino groups of lysine residues at theN-terminus of a histone. Unless otherwise indicated by context, the term“histone” is meant to refer to any histone protein, including H1, H2A,H2B, H3, H4, and H5, from any species. Preferred histone deacetylasesinclude class I and class II enzymes. Preferably the histone deacetylaseis a human HDAC, including, but not limited to, HDAC-1, HDAC-2, HDAC-3,HDAC-4, HDAC-5, HDAC-6, HDAC-7, HDAC-8, HDAC-9, HDAC-10, and HDAC-11. Insome other preferred embodiments, the histone deacetylase is derivedfrom a protozoal or fungal source. Preferred fungi include, but are notlimited to Saccharomyces cerevisiae, Candida spp. (such as C. albicans,C. glabrata, C. tropicalis, C. parapsilosis, C. krusei, C. lusitaniae,C. dubliniensis), Aspergillus spp. (such as A. fumigatus, A. flavus, A.niger, A. terreus), Fusarium spp., Paecilomyces lilacinus, Rhizopusarrhizus and Coccidioides immitis. In certain preferred embodiments, thehistone deacetylase is a fungal HDAC including, but not limited to Rpd3,Hos1, Hos2, Hda1, Hos3, Sir2, Hst, and homologs thereof. In preferredembodiments, a cleavage product of a prodrug compound of the presentinvention shows synergistic activity with an antifungal agent against afungal species, preferably at concentrations of inhibitor not toxic tomammalian cells. Preferably such antifungal agents are azole antifungalagents (a large number of active antifungal agents have an azolefunctionality as part of their structure; such an antifungal agent isgenerally referred to as an “antifungal azole”, an “azole antifungalagent” or an “azole”). Such combinations, and compositions thereof, canbe used to selectively treat fungal infection.

The term “antifungal agent” is intended to mean a substance capable ofinhibiting or preventing the growth, viability and/or reproduction of afungal cell. Preferable antifungal agents are those capable ofpreventing or treating a fungal infection in an animal or plant. Apreferable antifungal agent is a broad spectrum antifungal agent.However, an antifungal agent can also be specific to one or moreparticular species of fungus.

Preferred antifungal agents are ergosterol synthesis inhibitors, andinclude, but are not limited to azoles and phenpropimorph. Otherantifungal agents include, but are not limited to terbinafine. Preferredazoles include imidazoles and triazoles. Further preferred antifungalagents include, but are not limited to, ketoconazole, itroconazole,fluconazole, voriconazole, posaconazole, ravuconazole and miconazole.Like azoles, phenpropimorph is an ergosterol synthesis inhibitor, butacts on the ergosterol reductase (ERG24) step of the synthesis pathway.Terbinafine, is also an ergosterol inhibitor, but acts on the squaleneeposidase (ERG1) step.

The term “histone deacetylase inhibitor” or “inhibitor of histonedeacetylase” is used to identify a compound having a structure asdefined herein, which is capable of interacting with a histonedeacetylase and inhibiting its enzymatic activity. Inhibiting histonedeacetylase enzymatic activity means reducing the ability of a histonedeacetylase to remove an acetyl group from a histone. In some preferredembodiments, such reduction of histone deacetylase activity is at leastabout 50%, more preferably at least about 75%, and still more preferablyat least about 90%. In other preferred embodiments, histone deacetylaseactivity is reduced by at least 95% and more preferably by at least 99%.

Preferably, such inhibition is specific, i.e., the histone deacetylaseinhibitor reduces the ability of a histone deacetylase to remove anacetyl group from a histone at a concentration that is lower than theconcentration of the inhibitor that is required to produce another,unrelated biological effect. Preferably, the concentration of theinhibitor required for histone deacetylase inhibitory activity is atleast 2-fold lower, more preferably at least 5-fold lower, even morepreferably at least 10-fold lower, and most preferably at least 20-foldlower than the concentration required to produce an unrelated biologicaleffect. In certain preferred embodiments of the present invention,cleavage (e.g., hydrolysis) of the prodrug releases a compound (acleavage (e.g., hydrolyzation) product) which is an inhibitor of histonedeacetylase that is more active against a fungal histone deacetylasethan against a mammalian histone deacetylase. In certain preferredembodiments of the present invention, the inhibitor of histonedeacetylase is specific for a fungal histone deacetylase.

The terms “treating”, “treatment”, or the like, as used herein coversthe treatment of a disease-state in an animal and includes at least oneof: (i) preventing the disease-state from occurring, in particular, whensuch animal is predisposed to the disease-state but has not yetdeveloped symptoms of having it; (ii) inhibiting the disease-state,i.e., partially or completely arresting its development; (iii) relievingthe disease-state, i.e., causing regression of symptoms of thedisease-state, or ameliorating a symptom of the disease; and (iv)reversal or regression of the disease-state, preferably eliminating orcuring of the disease. In a preferred embodiment the terms “treating”,“treatment”, or the like, covers the treatment of a disease-state in ananimal and includes at least one of (ii), (iii) and (iv) above. In apreferred embodiment of the present invention the animal is a mammal,preferably a primate, more preferably a human. As is known in the art,adjustments for systemic versus localized delivery, age, body weight,general health, sex, diet, time of administration, drug interaction andthe severity of the condition may be necessary, and will beascertainable with routine experimentation by one of ordinary skill inthe art.

For simplicity, chemical moieties are defined and referred to throughoutprimarily as univalent chemical moieties (e.g., alkyl, aryl, etc.).Nevertheless, such terms are also used to convey correspondingmultivalent moieties under the appropriate structural circumstancesclear to those skilled in the art. For example, while an “alkyl” moietygenerally refers to a monovalent radical (e.g. CH₃—CH₂—), in certaincircumstances a bivalent linking moiety can be “alkyl,” in which casethose skilled in the art will understand the alkyl to be a divalentradical (e.g., —CH₂—CH₂—), which is equivalent to the term “alkylene.”(Similarly, in circumstances in which a divalent moiety is required andis stated as being “aryl,” those skilled in the art will understand thatthe term “aryl” refers to the corresponding divalent moiety, arylene).All atoms are understood to have their normal number of valences forbond formation (i.e., 4 for carbon, 3 for N, 2 for O, and 2, 4, or 6 forS, depending on the oxidation state of the S). On occasion a moiety maybe defined, for example, as (A)_(a)-B—, wherein a is 0 or 1. In suchinstances, when a is 0 the moiety is B- and when a is 1 the moiety isA-B—.

For simplicity, reference to a “C_(n)-C_(m),” heterocyclyl or“C_(n)-C_(m),” heteroaryl means a heterocyclyl or heteroaryl having from“n” to “m” annular atoms, where “n” and “m” are integers. Thus, forexample, a C₅-C₆-heterocyclyl is a 5- or 6-membered ring having at leastone heteroatom, and includes pyrrolidinyl (C₅) and piperidinyl (C₆);C₆-heteroaryl includes, for example, pyridyl and pyrimidyl.

The term “hydrocarbyl” refers to a straight, branched, or cyclic alkyl,alkenyl, or alkynyl, each as defined herein. A “C₀” hydrocarbyl is usedto refer to a covalent bond. Thus, “C₀-C₃-hydrocarbyl” includes acovalent bond, methyl, ethyl, ethenyl, ethynyl, propyl, propenyl,propynyl, and cyclopropyl.

The term “alkyl” is intended to mean a straight or branched chainaliphatic group having from 1 to 12 carbon atoms, preferably 1-8 carbonatoms, and more preferably 1-6 carbon atoms. Other preferred alkylgroups have from 2 to 12 carbon atoms, preferably 2-8 carbon atoms andmore preferably 2-6 carbon atoms. Preferred alkyl groups include,without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl,sec-butyl, tert-butyl, pentyl, and hexyl. A “C₀” alkyl (as in“C₀-C₃-alkyl”) is a covalent bond.

The term “alkenyl” is intended to mean an unsaturated straight orbranched chain aliphatic group with one or more carbon-carbon doublebonds, having from 2 to 12 carbon atoms, preferably 2-8 carbon atoms,and more preferably 2-6 carbon atoms. Preferred alkenyl groups include,without limitation, ethenyl, propenyl, butenyl, pentenyl, and hexenyl.

The term “alkynyl” is intended to mean an unsaturated straight orbranched chain aliphatic group with one or more carbon-carbon triplebonds, having from 2 to 12 carbon atoms, preferably 2-8 carbon atoms,and more preferably 2-6 carbon atoms. Preferred alkynyl groups include,without limitation, ethynyl, propynyl, butynyl, pentynyl, and hexynyl.

The terms “alkylene,” “alkenylene,” or “alkynylene” as used herein areintended to mean an alkyl, alkenyl, or alkynyl group, respectively, asdefined hereinabove, that is positioned between and serves to connecttwo other chemical groups. Preferred alkylene groups include, withoutlimitation, methylene, ethylene, propylene, and butylene. Preferredalkenylene groups include, without limitation, ethenylene, propenylene,and butenylene. Preferred alkynylene groups include, without limitation,ethynylene, propynylene, and butynylene.

The term “cycloalkyl” is intended to mean a saturated or unsaturatedmono-, bi, tri- or poly-cyclic hydrocarbon group having about 3 to 15carbons, preferably having 3 to 12 carbons, preferably 3 to 8 carbons,and more preferably 3 to 6 carbons. In certain preferred embodiments,the cycloalkyl group is fused to an aryl, heteroaryl or heterocyclicgroup. Preferred cycloalkyl groups include, without limitation,cyclopenten-2-enone, cyclopenten-2-enol, cyclohex-2-enone,cyclohex-2-enol, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl,cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

The term “heteroalkyl” is intended to mean a saturated or unsaturated,straight or branched chain aliphatic group, wherein one or more carbonatoms in the chain are independently replaced by a heteroatom selectedfrom the group consisting of O, S(O)₀₋₂, N and N(R³³).

The term “aryl” is intended to mean a mono-, bi-, tri- or polycyclicC₆-C₁₄ aromatic moiety, preferably comprising one to three aromaticrings. Preferably, the aryl group is a C₆-C₁₀ aryl group, morepreferably a C₆ aryl group. Preferred aryl groups include, withoutlimitation, phenyl, naphthyl, anthracenyl, and fluorenyl.

The terms “aralkyl” or “arylalkyl” is intended to mean a groupcomprising an aryl group covalently linked to an alkyl group. If anaralkyl group is described as “optionally substituted”, it is intendedthat either or both of the aryl and alkyl moieties may independently beoptionally substituted or unsubstituted. Preferably, the aralkyl groupis (C₁-C₆)alk(C₆-C₁₀)aryl, including, without limitation, benzyl,phenethyl, and naphthylmethyl. For simplicity, when written as“arylalkyl” this term, and terms related thereto, is intended toindicate the order of groups in a compound as “aryl-alkyl”. Similarly,“alkyl-aryl” is intended to indicate the order of the groups in acompound as “alkyl-aryl”.

The terms “heterocyclyl”, “heterocyclic” or “heterocycle” are intendedto mean a group which is a mono-, bi-, or polycyclic structure havingfrom about 3 to about 14 atoms, wherein one or more atoms areindependently selected from the group consisting of N, O, and S. Thering structure may be saturated, unsaturated or partially unsaturated.In certain preferred embodiments, the heterocyclic group isnon-aromatic. In a bicyclic or polycyclic structure, one or more ringsmay be aromatic; for example one ring of a bicyclic heterocycle or oneor two rings of a tricyclic heterocycle may be aromatic, as in indan and9,10-dihydro anthracene. Preferred heterocyclic groups include, withoutlimitation, epoxy, aziridinyl, tetrahydrofuranyl, pyrrolidinyl,piperidinyl, piperazinyl, thiazolidinyl, oxazolidinyl, oxazolidinonyl,and morpholino. In certain preferred embodiments, the heterocyclic groupis fused to an aryl, heteroaryl, or cycloalkyl group. Examples of suchfused heterocycles include, without limitation, tetrahydroquinoline anddihydrobenzofuran. Specifically excluded from the scope of this term arecompounds where an annular O or S atom is adjacent to another O or Satom.

In certain preferred embodiments, the heterocyclic group is a heteroarylgroup. As used herein, the term “heteroaryl” is intended to mean amono-, bi-, tri- or polycyclic group having 5 to 14 ring atoms,preferably 5, 6, 9, or 10 ring atoms; having 6, 10, or 14 pi electronsshared in a cyclic array; and having, in addition to carbon atoms,between one or more heteroatoms independently selected from the groupconsisting of N, O, and S. For example, a heteroaryl group may bepyrimidinyl, pyridinyl, benzimidazolyl, thienyl, benzothiazolyl,benzofuranyl and indolinyl. Preferred heteroaryl groups include, withoutlimitation, thienyl, benzothienyl, furyl, benzofuryl, dibenzofuryl,pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl,indolyl, quinolyl, isoquinolyl, quinoxalinyl, tetrazolyl, oxazolyl,thiazolyl, and isoxazolyl.

The terms “arylene,” “heteroarylene,” or “heterocyclylene” are intendedto mean an aryl, heteroaryl, or heterocyclyl group, respectively, asdefined hereinabove, that is positioned between and serves to connecttwo other chemical groups.

Preferred heterocyclyls and heteroaryls include, but are not limited to,acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl,benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl,benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl,carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl,cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furyl, furazanyl,imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl,indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl,isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl,isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl,naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl,1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl,oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl,phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl,piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl,pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl,pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole,pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl,pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl,quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl,tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, thiadiazolyl(e.g., 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl,1,3,4-thiadiazolyl), thianthrenyl, thiazolyl, thienyl, thienothiazolyl,thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl, triazolyl(e.g., 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl,1,3,4-triazolyl), and xanthenyl.

Aromatic polycycles include, but are not limited to, bicyclic andtricyclic fused ring systems, including for example naphthyl.

Non-aromatic polycycles include, but are not limited to, bicyclic andtricyclic fused ring systems where each ring can be 4-9 membered andeach ring can containing zero, 1 or more double and/or triple bonds.Suitable examples of non-aromatic polycycles include, but are notlimited to, decalin, octahydroindene, perhydrobenzocycloheptene andperhydrobenzo-M-azulene.

Polyheteroaryl groups include bicyclic and tricyclic fused rings systemswhere each ring can independently be 5 or 6 membered and contain one ormore heteroatom, for example, 1, 2, 3 or 4 heteroatoms, independentlychosen from O, N and S such that the fused ring system is aromatic.Suitable examples of polyheteroaryl ring systems include quinoline,isoquinoline, pyridopyrazine, pyrrolopyridine, furopyridine, indole,benzofuran, benzothiofuran, benzindole, benzoxazole, pyrroloquinoline,and the like.

Non-aromatic polyheterocyclic groups include but are not limited tobicyclic and tricyclic ring systems where each ring can be 4-9 membered,contain one or more heteratom, for example 1, 2, 3 or 4 heteratoms,independently chosen from O, N and S, and contain zero, or one or moreC—C double or triple bonds. Suitable examples of non-aromaticpolyheterocycles include but are not limited to, hexitol,cis-perhydro-cyclohepta[b]pyridinyl, decahydro-benzo[f][1,4]oxazepinyl,2,8-dioxabicyclo[3.3.0]octane, hexahydro-thieno[3,2-b]thiophene,perhydropyrrolo[3,2-b]pyrrole, perhydronaphthyridine,perhydrop-1H-dicyclopenta[b,e]pyran.

Mixed aryl and non-aryl polyheterocycle groups include but are notlimited to bicyclic and tricyclic fused ring systems where each ring canbe 4-9 membered, contain one or more heteroatom independently chosenfrom O, N and S and at least one of the rings must be aromatic. Suitableexamples of mixed aryl and non-aryl polyheteorcycles include2,3-dihydroindole, 1,2,3,4-tetrahydroquinoline,5,11-dihydro-10H-dibenz[b,e][1,4]diazepine,5H-dibenzo[b,e][1,4]diazepine,1,2-dihydropyrrolo[3,4-b][1,5]benzodiazepine,1,5-dihydropyrido[2,3-b][1,4]diazepin-4-one,1,2,3,4,6,11-hexhydro-benzo[b]pyrido[2,3-e][1,4]diazepine-5-one,methylenedioxyphenyl, bis-methylenedioxyphenyl,1,2,3,4-tetrahydronaphthalene, dibenzosuberane dihydroanthracene and9H-fluorene.

As employed herein, and unless stated otherwise, when a moiety (e.g.,alkyl, heteroalkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, etc.) isdescribed as “optionally substituted” it is meant that the groupoptionally has from one to four, preferably from one to three, morepreferably one or two, non-hydrogen substituents. Suitable substituentsinclude, without limitation, halo, hydroxy, oxo (e.g., an annular —CH—substituted with oxo is —C(O)—) nitro, halohydrocarbyl, hydrocarbyl,alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, aralkyl, alkoxy,aryloxy, amino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl,acyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl,alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl,acyloxy, cyano, and ureido groups. Preferred substituents, which arethemselves not further substituted (unless expressly stated otherwise)are:

-   -   (a) halo, cyano, oxo, carboxy, formyl, nitro, amino, amidino,        guanidino,    -   (b) C₁-C₅ alkyl or alkenyl or arylalkyl imino, carbamoyl, azido,        carboxamido, mercapto, hydroxy, hydroxyalkyl, alkylaryl,        arylalkyl, C₁-C₈ alkyl, C₁-C₈ alkenyl, C₁-C₈ alkoxy, C₁-C₈        alkoxycarbonyl, aryloxycarbonyl, C₂-C₈ acyl, C₂-C₈ acylamino,        C₁-C₈ alkylthio, arylalkylthio, arylthio, C₁-C₈ alkylsulfinyl,        arylalkylsulfinyl, arylsulfinyl, C₁-C₈ alkylsulfonyl,        arylalkylsulfonyl, arylsulfonyl, C₀-C₆ N-alkyl carbamoyl, C₂-C₁₅        N,N-dialkylcarbamoyl, C₃-C₇ cycloalkyl, aroyl, aryloxy,        arylalkyl ether, aryl, aryl fused to a cycloalkyl or heterocycle        or another aryl ring, C₃-C₇ heterocycle, C₅-C₁₅ heteroaryl or        any of these rings fused or spiro-fused to a cycloalkyl,        heterocyclyl, or aryl, wherein each of the foregoing is further        optionally substituted with one more moieties listed in (a),        above; and    -   (c) —(CR³²R^(33a))_(s)—NR³⁰R³¹, wherein s is from 0 (in which        case the nitrogen is directly bonded to the moiety that is        substituted) to 6, R³² and R^(33a) are each independently        hydrogen, halo, hydroxyl or C₁-C₄alkyl, and R³⁰ and R³¹ are each        independently hydrogen, cyano, oxo, hydroxyl, —C₁-C₈ alkyl,        C₁-C₈ heteroalkyl, C₁-C₈ alkenyl, carboxamido, C₁-C₃        alkyl-carboxamido, carboxamido-C₁-C₃ alkyl, amidino,        C₂-C₈hydroxyalkyl, C₁-C₃ alkylaryl, aryl-C₁-C₃ alkyl, C₁-C₃        alkylheteroaryl, heteroaryl-C₁-C₃ alkyl, C₁-C₃        alkylheterocyclyl, heterocyclyl-C₁-C₃ alkyl C₁-C₃        alkylcycloalkyl, cycloalkyl-C₁-C₃ alkyl, C₂-C₈ alkoxy, C₂-C₈        alkoxy-C₁-C₄alkyl, C₁-C₈ alkoxycarbonyl, aryloxycarbonyl,        aryl-C₁-C₃ alkoxycarbonyl, heteroaryloxycarbonyl,        heteroaryl-C₁-C₃ alkoxycarbonyl, C₁-C₈ acyl, C₀-C₈        alkyl-carbonyl, aryl-C₀-C₈ alkyl-carbonyl, heteroaryl-C₀-C₈        alkyl-carbonyl, cycloalkyl-C₀-C₈ alkyl-carbonyl, C₀-C₈        alkyl-NH-carbonyl, aryl-C₀-C₈ alkyl-NH-carbonyl,        heteroaryl-C₀-C₈ alkyl-NH-carbonyl, cycloalkyl-C₀-C₈        alkyl-NH-carbonyl, C₀-C₈ alkyl-O-carbonyl, aryl-C₀-C₈        alkyl-O-carbonyl, heteroaryl-C₀-C₈ alkyl-O-carbonyl,        cycloalkyl-C₀-C₈ alkyl-O-carbonyl, C₁-C₈ alkylsulfonyl,        arylalkylsulfonyl, arylsulfonyl, heteroarylalkylsulfonyl,        heteroarylsulfonyl, C₁-C₈ alkyl-NH-sulfonyl,        arylalkyl-NH-sulfonyl, aryl-NH-sulfonyl,        heteroarylalkyl-NH-sulfonyl, heteroaryl-NH-sulfonyl aroyl, aryl,        cycloalkyl, heterocyclyl, heteroaryl, aryl-C₁-C₃ alkyl-,        cycloalkyl-C₁-C₃ alkyl-, heterocyclyl-C₁-C₃ alkyl-,        heteroaryl-C₁-C₃ alkyl-, or protecting group, wherein each of        the foregoing is further optionally substituted with one more        moieties listed in (a), above; or

R³⁰ and R³¹ taken together with the N to which they are attached form aheterocyclyl or heteroaryl, each of which is optionally substituted withfrom 1 to 3 substituents selected from the group consisting of (a)above, a protecting group, and (X³⁰—Y³¹—), wherein said heterocyclyl mayalso be bridged (forming a bicyclic moiety with a methylene, ethylene orpropylene bridge); wherein

X³⁰ is selected from the group consisting of C₁-C₈alkyl, C₂-C₈alkenyl-,C₂-C₈alkynyl-, —C₀-C₃alkyl-C₂-C₈alkenyl-C₀-C₃alkyl,C₀-C₃alkyl-C₂-C₈alkynyl-C₀-C₃alkyl, C₀-C₃alkyl-O—C₀-C₃alkyl-,HO—C₀-C₃alkyl-, C₀-C₄alkyl-N(R³⁰)—C₀-C₃alkyl-, N(R³⁰)(R³¹)—C₀-C₃alkyl-,N(R³⁰)(R³¹)—C₀-C₃alkenyl-, N(R³⁰)(R³¹)—C₀-C₃alkynyl-,(N(R³⁰)(R³¹))₂—C═N—, C₀-C₃alkyl-S(O)₀₋₂—C₀-C₃alkyl-, CF₃—C₀-C₃alkyl-,C₁-C₈heteroalkyl, aryl, cycloalkyl, heterocyclyl, heteroaryl,aryl-C₁-C₃alkyl-, cycloalkyl-C₁-C₃alkyl-, heterocyclyl-C₁-C₃alkyl-,heteroaryl-C₁-C₃alkyl-, N(R³⁰)(R³¹)-heterocyclyl-C₁-C₃alkyl-, whereinthe aryl, cycloalkyl, heteroaryl and heterocycyl are optionallysubstituted with from 1 to 3 substituents from (a); and Y³¹ is selectedfrom the group consisting of a direct bond, —O—, —N(R³⁰)—, —C(O)—,—O—C(O)—, —C(O)—O—, —N(R³⁰)—C(O)—, —C(O)—N(R³⁰)—, —N(R³⁰)—C(S)—,—C(S)—N(R³⁰)—, —N(R³⁰)—C(O)—N(R³¹)—, —N(R³⁰)—C(NR³⁰)—N(R³¹)—,—N(R³⁰)—C(NR³¹)—, —C(NR³¹)—N(R³⁰), —N(R³⁰)—C(S)—N(R³¹)—,—N(R³⁰)—C(O)—O—, —O—C(O)—N(R³¹)—, —N(R³⁰)—C(S)—O—, —O—C(S)—N(R³¹)—,—S(O)₀₋₂—, —SO₂N(R³¹)—, —N(R³¹)—SO₂— and —N(R³⁰)—SO₂N(R³¹)—.

As a non-limiting example, substituted phenyls include 2-fluorophenyl,3,4-dichlorophenyl, 3-chloro-4-fluoro-phenyl, 2-fluoro-3-propylphenyl.As another non-limiting example, substituted n-octyls include2,4-dimethyl-5-ethyl-octyl and 3-cyclopentyl-octyl. Included within thisdefinition are methylenes (—CH₂—) substituted with oxygen to formcarbonyl —CO—.

When there are two optional substituents bonded to adjacent atoms of aring structure, such as for example phenyl, thiophenyl, or pyridinyl,the substituents, together with the atoms to which they are bonded,optionally form a 5- or 6-membered cycloalkyl or heterocycle having 1,2, or 3 annular heteroatoms.

In a preferred embodiment, hydrocarbyl, alkyl, alkenyl, alkynyl,heteroalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, aromaticpolycycle, non-aromatic polycycle, polyheteroaryl, non-aromaticpolyheterocyclic and mixed aryl and non-aryl polyheterocycle groups areunsubstituted.

In other preferred embodiments, hydrocarbyl, alkyl, alkenyl, alkynyl,heteroalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, aromaticpolycycle, non-aromatic polycycle, polyheteroaryl, non-aromaticpolyheterocyclic and mixed aryl and non-aryl polyheterocycle groups aresubstituted with from 1 to 3 independently selected substituents.

Preferred substituents on alkyl groups include, but are not limited to,hydroxyl, halogen (e.g., a single halogen substituent or multiple halosubstituents; in the latter case, groups such as CF₃ or an alkyl groupbearing more than one Cl), cyano, nitro, alkyl, cycloalkyl, alkenyl,cycloalkenyl, alkynyl, heterocycle, aryl, —OR^(u), —SR^(u), —S(═O)R^(y),—S(═O)₂R^(y), —P(═O)₂R^(y), —S(═O)₂OR^(y), —P(═O)₂OR^(y), —NR^(v)R^(w),—NR^(v)S(═O)₂R^(y), —NR^(v)P(═O)₂R^(y), —S(═O)₂NR^(v)R^(w),—P(═O)₂NR^(v)R^(w), —C(═O)OR^(y), —C(═O)R^(u), —C(═O)NR^(v)R^(w),—OC(═O)R^(u), —OC(═O)NR^(v)R^(w), —NR^(v)C(═O)OR^(y),—NR^(xx)C(═O)NR^(v)R^(w), —NR^(xx)S(═O)₂NR^(v)R^(w),—NR^(xx)P(═O)₂NR^(v)R^(w), —NR^(v)C(═O)R^(u) or —NR^(v)P(═O)₂R^(y),wherein R^(u) is hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkynyl, heterocycle or aryl; R^(v), R^(w) and R^(xx) are independentlyhydrogen, alkyl, cycloalkyl, heterocycle or aryl, or said R^(v) andR^(w) together with the N to which they are bonded optionally form aheterocycle; and R^(y) is alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkynyl, heterocycle or aryl. In the aforementioned exemplarysubstituents, groups such as alkyl, cycloalkyl, alkenyl, alkynyl,cycloalkenyl, heterocycle and aryl can themselves be optionallysubstituted.

Preferred substituents on alkenyl and alkynyl groups include, but arenot limited to, alkyl or substituted alkyl, as well as those groupsrecited as preferred alkyl substituents.

Preferred substituents on cycloalkyl groups include, but are not limitedto, nitro, cyano, alkyl or substituted alkyl, as well as those groupsrecited about as preferred alkyl substituents. Other preferredsubstituents include, but are not limited to, spiro-attached or fusedcyclic substituents, preferably spiro-attached cycloalkyl,spiro-attached cycloalkenyl, spiro-attached heterocycle (excludingheteroaryl), fused cycloalkyl, fused cycloalkenyl, fused heterocycle, orfused aryl, where the aforementioned cycloalkyl, cycloalkenyl,heterocycle and aryl substituents can themselves be optionallysubstituted.

Preferred substituents on cycloalkenyl groups include, but are notlimited to, nitro, cyano, alkyl or substituted alkyl, as well as thosegroups recited as preferred alkyl substituents. Other preferredsubstituents include, but are not limited to, spiro-attached or fusedcyclic substituents, especially spiro-attached cycloalkyl,spiro-attached cycloalkenyl, spiro-attached heterocycle (excludingheteroaryl), fused cycloalkyl, fused cycloalkenyl, fused heterocycle, orfused aryl, where the aforementioned cycloalkyl, cycloalkenyl,heterocycle and aryl substituents can themselves be optionallysubstituted.

Preferred substituents on aryl groups include, but are not limited to,nitro, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substitutedcycloalkenyl, cyano, alkyl or substituted alkyl, as well as those groupsrecited above as preferred alkyl substituents. Other preferredsubstituents include, but are not limited to, fused cyclic groups,especially fused cycloalkyl, fused cycloalkenyl, fused heterocycle, orfused aryl, where the aforementioned cycloalkyl, cylcoalkenyl,heterocycle and aryl substituents can themselves be optionallysubstituted. Still other preferred substituents on aryl groups (phenyl,as a non-limiting example) include, but are not limited to, haloalkyland those groups recited as preferred alkyl substituents.

Preferred substituents on heterocylic groups include, but are notlimited to, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, nitro, oxo (i.e., ═O), cyano, alkyl,substituted alkyl, as well as those groups recited as preferred alkylsubstituents. Other preferred substituents on heterocyclic groupsinclude, but are not limited to, spiro-attached or fused cylicsubstituents at any available point or points of attachment, morepreferably spiro-attached cycloalkyl, spiro-attached cycloalkenyl,spiro-attached heterocycle (excluding heteroaryl), fused cycloalkyl,fused cycloakenyl, fused heterocycle and fused aryl, where theaforementioned cycloalkyl, cycloalkenyl, heterocycle and arylsubstituents can themselves be optionally substituted.

In a preferred embodiment, a heterocyclic group is substituted oncarbon, nitrogen and/or sulfur at one or more positions. Preferredsubstituents on nitrogen include, but are not limited to N-oxide, alkyl,aryl, aralkyl, alkylcarbonyl, alkylsulfonyl, arylcarbonyl, arylsulfonyl,alkoxycarbonyl, or aralkoxycarbonyl. Preferred substituents on sulfurinclude, but are not limited to, oxo and C₁₋₆alkyl. In certain preferredembodiments, nitrogen and sulfur heteroatoms may independently beoptionally oxidized and nitrogen heteroatoms may independently beoptionally quaternized.

Especially preferred substituents on alkyl groups include halogen andhydroxy.

Especially preferred substituents on ring groups, such as aryl,heteroaryl, cycloalkyl and heterocyclyl, include halogen, alkoxy andalkyl.

Preferred substituents on aromatic polycycles include, but are notlimited to, oxo, C₁-C₆alkyl, cycloalkylalkyl (e.g. cyclopropylmethyl),oxyalkyl, halo, nitro, amino, alkylamino, aminoalkyl, alkyl ketones,nitrile, carboxyalkyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl andOR^(aa), such as alkoxy, wherein R^(aa) is selected from the groupconsisting of H, C₁-C₆alkyl, C₄-C₉cycloalkyl, C₄-C₉heterocycloalkyl,aryl, heteroaryl, arylalkyl, heteroarylalkyl and (CH₂)₀₋₆Z^(a)R^(bb),wherein Z^(a) is selected from the group consisting of O, NR^(cc), S andS(O), and R^(bb) is selected from the group consisting of H, C₁-C₆alkyl,C₄-C₉cycloalkyl, C₄-C₉heterocycloalkyl, C₄-C₉heterocycloalkylalkyl,aryl, mixed aryl and non-aryl polycycle, heteroaryl, arylalkyl, (e.g.benzyl), and heteroarylalkyl (e.g. pyridylmethyl); and R^(cc) isselected from the group consisting of H, C₁-C₆alkyl, C₄-C₉cycloalkyl,C₄-C₉heterocycloalkyl, aryl, heteroaryl, arylalkyl (e.g. benzyl),heteroarylalkyl (e.g. pyridylmethyl) and amino acyl.

Preferred substituents on non-aromatic polycycles include, but are notlimited to, oxo, C₃-C₉cycloalkyl, such as cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl and the like. Unless otherwise noted,non-aromatic polycycle substituents include both unsubstitutedcycloalkyl groups and cycloalkyl groups that are substituted by one ormore suitable substituents, including but not limited to, C₁-C₆alkyl,oxo, halo, hydroxy, aminoalkyl, oxyalkyl, alkylamino and OR^(aa), suchas alkoxy. Preferred substituents for such cycloalkyl groups includehalo, hydroxy, alkoxy, oxyalkyl, alkylamino and aminoalkyl.

Preferred substituents on carbon atoms of polyheteroaryl groups includebut are not limited to, straight and branched optionally substitutedC₁-C₆alkyl, unsaturation (i.e., there are one or more double or tripleC—C bonds), acyl, oxo, cycloalkyl, halo, oxyalkyl, alkylamino,aminoalkyl, acylamino, OR^(aa) (for example alkoxy), and a substituentof the formula —O—(CH₂CH═CH(CH₃)(CH₂))₁₋₃H. Examples of suitablestraight and branched C₁-C₆alkyl substituents include but are notlimited to methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl,t-butyl and the like. Preferred substituents include halo, hydroxy,alkoxy, oxyalkyl, alkylamino and aminoalkyl. Preferably substitutions onnitrogen atoms include, for example by N-oxide or R^(cc). Preferredsubstituents on nitrogen atoms include H, C₁-C₄alkyl, acyl, aminoacyland sulfonyl. Preferably sulfur atoms are unsubstituted. Preferredsubstituents on sulfur atoms include but are not limited to oxo andlower alkyl.

Preferred substituents on carbon atoms of non-aromatic polyheterocyclicgroups include but are not limited to straight and branched optionallysubstituted C₁-C₆alkyl, unsaturation (i.e., there are one or more doubleor triple C—C bonds), acyl, oxo, cycloalkyl, halo, oxyalkyl, alkylamino,aminoalkyl, acylamino and OR^(aa), for example alkoxy. Examples ofsuitable straight and branched C₁-C₆alkyl substituents include but arenot limited to methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl,t-butyl and the like. Preferred substituents include halo, hydroxy,alkoxy, oxyalkyl, alkylamino and aminoalkyl. Preferably substitutions onnitrogen atoms include, for example, N-oxide or R^(cc). Preferred Nsubstituents include H, C₁-C₄ alkyl, acyl, aminoacyl and sulfonyl.Preferably, sulfur atoms are unsubstituted. Preferred S substituentsinclude oxo and lower alkyl.

Preferred substituents on mixed aryl and non-aryl polyheterocycle groupsinclude, but are not limited to, nitro or as described above fornon-aromatic polycycle groups. Preferred substituents on carbon atomsinclude, but are not limited to, —N—OH, ═N—OH, optionally substitutedalkyl, unsaturation (i.e., there are one or more double or triple C—Cbonds), oxo, acyl, cycloalkyl, halo, oxyalkyl, alkylamino, aminoalkyl,acylamino and OR^(aa), for example alkoxy. Preferably substitutions onnitrogen atoms include, for example, N-oxide or R^(cc). Preferred Nsubstituents include H, C₁₋₄alkyl, acyl aminoacyl and sulfonyl.Preferably sulfur atoms are unsubstituted. Preferred S substituentsinclude oxo and lower alkyl.

A “halohydrocarbyl” is a hydrocarbyl moiety in which from one to allhydrogens have been replaced with one or more halo.

The term “halogen” or “halo” is intended to mean chlorine, bromine,fluorine, or iodine. As herein employed, the term “acyl” refers to analkylcarbonyl or arylcarbonyl substituent. The term “acylamino” refersto an amide group attached at the nitrogen atom (i.e., R—CO—NH—). Theterm “carbamoyl” refers to an amide group attached at the carbonylcarbon atom (i.e., NH₂—CO—). The nitrogen atom of an acylamino orcarbamoyl substituent is additionally optionally substituted. The term“sulfonamido” refers to a sulfonamide substituent attached by either thesulfur or the nitrogen atom. The term “amino” is meant to include NH₂,alkylamino, arylamino, and cyclic amino groups. The term “ureido” asemployed herein refers to a substituted or unsubstituted urea moiety.

The term “radical” is intended to mean a chemical moiety comprising oneor more unpaired electrons.

Where optional substituents are chosen from “one or more” groups it isto be understood that this definition includes all substituents beingchosen from one of the specified groups or the substituents being chosenfrom two or more of the specified groups.

In addition, substituents on cyclic moieties (i.e., cycloalkyl,heterocyclyl, aryl, heteroaryl) include 5-6 membered mono- and 9-14membered bi-cyclic moieties fused to the parent cyclic moiety to form abi- or tri-cyclic fused ring system. Substituents on cyclic moietiesalso include 5-6 membered mono- and 9-14 membered bi-cyclic moietiesattached to the parent cyclic moiety by a covalent bond to form a bi- ortri-cyclic bi-ring system. For example, an optionally substituted phenylincludes, but is not limited to, the following:

An “unsubstituted” moiety (e.g., unsubstituted cycloalkyl, unsubstitutedheteroaryl, etc.) means that moiety as defined above that does not havean optional substituent. Thus, for example, “unsubstituted aryl” doesnot include phenyl substituted with a halo.

As used herein, “an amino protecting group” refers to any functionalgroup commonly used to protect an α-amino group. Suitable aminoprotecting groups include, but are not limited to, t-butyloxycarbonyl,isoamyloxycarbonyl, o-nitrophenylsulfenyl, fluoroenylmethyloxycarbonyl,o-nitropyridinylsulfenyl and biphenylproploxycarbonyl.

An “amino acid residue” refers to any residue of a natural or unnaturalamino acid, non-limiting examples of which are residues of alanine,arginine, asparagine, aspartic acid, cysteine, homocysteine, glutamine,glutamic acid, isoleucine, norleucine, glycine, phenylglycine, leucine,histidine, methionine, lysine, phenylalanine, homophenylalanine,ornithine, praline, serine, homoserine, valine, norvaline, threonine,tryptophane, tyrosine and the like. With the exception of glycine, allamino acids may be in the D-, L- or D,L-form.

The term “radical” is intended to mean a chemical moiety comprising oneor more unpaired electrons.

Some compounds of the invention may have one or more chiral centersand/or geometric isomeric centers (E- and Z-isomers), and it is to beunderstood that the invention encompasses all such optical,diastereoisomers and geometric isomers. The invention also comprises alltautomeric forms of the compounds disclosed herein.

All of the compounds in this application were named using Chemdraw Ultraversion 9 or 10, which are available through Cambridgesoft.co, 100Cambridge Park Drive, Cambridge, Mass. 02140.

Compounds

In a first aspect, the invention provides prodrugs of inhibitors ofhistone deacetylase, the prodrugs having the formula (1):

Cy-L¹-Ar-Y¹—C(O)—N(R^(x))—Z  (1)

and pharmaceutically acceptable salts thereof, wherein

Cy is —H, cycloalkyl, aryl, heteroaryl, or heterocyclyl, any of whichmay be optionally substituted;

L¹ is —(CH₂)_(m)—W—, where m is 0, 1, 2, 3, or 4, and W is selected fromthe group consisting of —C(O)NH—, —S(O)₂NH—, —NHC(O)—, —NHS(O)₂—, and—NH—C(O)—NH—;

Ar is arylene, wherein said arylene optionally may be additionallysubstituted and optionally may be fused to an aryl or heteroaryl ring,or to a saturated or partially unsaturated cycloalkyl or heterocyclicring, any of which may be optionally substituted;

Y¹ is a chemical bond or a straight- or branched-chain saturatedalkylene, wherein said alkylene may be optionally substituted;

R^(x) is H or —OH;

-   Z is —R²⁰, —O—R²⁰, —R²¹, or

wherein —R²⁰ is selected from the group consisting of —C(O)—R¹⁰,—C(O)O—R¹⁰, —R¹¹, —CH(R¹²)—O—C(O)—R¹⁰,—C(O)—C[(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³),—S(O₂)R¹⁰—C(O)—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰,—C(O)—(CH₂)₁₋₄—C(OH)(COOR¹⁰)—(CH₂)₁₋₄—COOR¹⁰,—C(O)-[C(R¹⁴)(R¹⁴)]₁₄—P(O)(OH)(OH),—C(O)—(CH₂)₁₋₄—N(R¹⁴)—C[═N(R^(10′))]-N(R^(10′))(R^(10′)),—C(O)—(CH₂)—CH(OH)—(CH₂)—N(CH₃)(CH₃), —C(O)—CH(NH₂)—(CH₂)₁₋₆—COOH(preferably —C(O)—CH(NH₂)—(CH₂)—COOH),—C(O)—O—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰ and —C(O)—(CH₂)_(n)—C(O)OR¹⁰,provided that the N to which Z is bound is not directly bonded to two Oatoms and further provided that (a) when Z is —R²⁰ then R^(x) is —OH,and (b) when Z is —OR²⁰ then R^(x) is —H; or

R^(x) is absent and R²⁰ forms an optionally substituted heterocyclicring with the N to which it is attached;

n is 0, 1, 2, 3, or 4, preferably 1, 2, 3, or 4;

each R¹⁰ is independently selected from the group consisting ofhydrogen, optionally substituted C₁-C₂₀ alkyl, optionally substitutedC₂-C₂₀ alkenyl, optionally substituted C₂-C₂₀ alkynyl, optionallysubstituted C₁-C₂₀ alkoxycarbonyl, optionally substituted cycloalkyl,optionally substituted heterocycloalkyl, optionally substituted aryl,optionally substituted heteroaryl, optionally substitutedcycloalkylalkyl, optionally substituted heterocycloalkylalkyl,optionally substituted arylalkyl, optionally substitutedheteroarylalkyl, optionally substituted cycloalkylalkenyl, optionallysubstituted heterocycloalkylalkenyl, optionally substituted arylalkenyl,optionally substituted heteroarylalkenyl, optionally substitutedcycloalkylalkynyl, optionally substituted heterocycloalkylalkynyl,optionally substituted arylalkynyl, optionally substitutedheteroarylalkynyl, optionally substituted alkylcycloalkyl, optionallysubstituted alkylheterocycloalkyl, optionally substituted alkylaryl,optionally substituted alkylheteroaryl, optionally substitutedalkenylcycloalkyl, optionally substituted alkenylheterocycloalkyl,optionally substituted alkenylaryl, optionally substitutedalkenylheteroaryl, optionally substituted alkynylcycloalkyl, optionallysubstituted alkynylheterocycloalkyl, optionally substituted alkynylary,optionally substituted alkynylheteroaryl, a sugar residue, and an aminoacid residue (preferably bonded through the carboxy terminus of theamino acid);

each R^(10′) is independently hydrogen or C₁₋₆alkyl, or

R¹⁰ and R^(10′) together with the carbon atom to which they are attachedform an optionally substituted spirocycloalkyl;

R²¹ is a sugar or -amino acid-R¹³, wherein R¹³ is covalently bound tothe N-terminus;

R¹¹ is selected from the group consisting of hydrogen, optionallysubstituted heterocycloalkyl, optionally substituted aryl, andoptionally substituted heteroaryl;

R¹² is selected from hydrogen or alkyl; and

R¹³ is selected from the group consisting of hydrogen,—C(O)—CH[N(R^(10′))(R^(10′))]—C₁-C₆alkyl,—C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-N(R^(10′))(R^(10′)),—C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-aryl,—C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-heteroaryl, —C(O)-aryl,—C(O)-heteroaryl, an amino protecting group, and R¹⁰; and

each R¹⁴ is independently selected from the group consisting of H,C₁-C₆alkyl and cycloalkyl, or two R¹⁴, together with the atom to whichthey are attached, form a cycloalkyl.

In certain preferred embodiments, Cy is C₆-C₁₄ aryl, more preferablyC₆-C₁₀ aryl, and most preferably phenyl or naphthyl, any of which may beoptionally substituted. In certain other preferred embodiments, Cy isheteroaryl. In some preferred embodiments, the heteroaryl group isselected from the group consisting of thienyl, benzothienyl, furyl,benzofuryl, quinolyl, isoquinolyl, and thiazolyl, any of which may beoptionally substituted. In certain particularly preferred embodiments,Cy is selected from the group consisting of phenyl, naphthyl, thienyl,benzothienyl, and quinolyl, any of which may be optionally substituted.In certain other preferred embodiments, Cy is phenyl, pyridine orindole, more preferably phenyl or indole. In certain preferredembodiments, Cy is substituted with one or more substituents selectedfrom the group consisting of trihaloalkyl (preferably trifluoroalkyl),halogen, CN, amidine, sulfone, alkylsulfone, imidate and alkylimidate.In certain preferred embodiments, Cy is phenyl substituted with one ormore substituents selected from the group consisting of trihaloalkyl(preferably trifluoroalkyl), halogen, CN, amidine, sulfone,alkylsulfone, imidate and alkylimidate, preferably selected from thegroup consisting of trihaloalkyl (preferably trifluoroalkyl) andhalogen.

L¹ is —(CH₂)_(m)—W—, where m is 0, 1, 2, 3, or 4, and W is selected fromthe group consisting of —C(O)NH—, —S(O)₂NH—, —NHC(O)—, —NHS(O)₂—, and—NH—C(O)—NH—. Preferably, m is 0, 1, or 2, more preferably 0 or 1.

Preferably, Ar is C₆-C₁₄ arylene, more preferably C₆-C₁₀ arylene, any ofwhich may be additionally substituted. In certain preferred embodiments,Ar is phenylene, preferably 4-phenylene. In some preferred embodiments,the phenylene is fused to an aryl or heteroaryl ring, or to a saturatedor partially unsaturated cycloalkyl or heterocyclic ring, any of whichgroups also may be optionally substituted.

Y¹ is a chemical bond or is a straight- or branched-chain alkylene,which may be optionally substituted. In some preferred embodiments, Y¹is a chemical bond, and the group —C(O)NH—Z is directly attached to Ar.In some other preferred embodiments, Y¹ is alkylene, preferablysaturated alkylene. Preferably, the saturated alkylene is C₁-C₈alkylene, more preferably C₁-C₆ alkylene, still more preferably C₁-C₃alkylene, and yet still more preferably C₁-C₂ alkylene, any of which maybe optionally substituted. In some particularly preferred embodiments,Y¹ is methylene.

Substituted alkyl, aryl, heterocyclyl, and heteroaryl groups have one ormore, preferably between one and about three, more preferably one or twosubstituents, which are preferably selected from the group consisting ofC₁-C₆ alkyl, preferably C₁-C₄ alkyl; halo, preferably Cl, Br, or F;haloalkyl, preferably (halo)₁₋₅(C₁-C₆)alkyl, more preferably(halo)₁₋₅(C₁-C₃)alkyl, and most preferably CF₃; C₁-C₆ alkoxy, preferablymethoxy, ethoxy, or benzyloxy; C₆-C₁₀ aryloxy, preferably phenoxy; C₁-C₆alkoxycarbonyl, preferably C₁-C₃ alkoxycarbonyl, most preferablycarbomethoxy or carboethoxy; C₆-C₁₀ aryl, preferably phenyl;(C₆-C₁₀)ar(C₁-C₆)alkyl, preferably (C₆-C₁₀)ar(C₁-C₃)alkyl, morepreferably benzyl, naphthylmethyl or phenethyl; hydroxy(C₁-C₆)alkyl,preferably hydroxy(C₁-C₃)alkyl, more preferably hydroxymethyl;amino(C₁-C₆)alkyl, preferably amino(C₁-C₃)alkyl, more preferablyaminomethyl; (C₁-C₆)alkylamino, preferably methylamino, ethylamino, orpropylamino; di-(C₁-C₆)alkylamino, preferably dimethylamino ordiethylamino; (C₁-C₆)alkylcarbamoyl, preferably methylcarbamoyl,dimethylcarbamoyl, or benzylcarbamoyl; (C₆-C₁₀)arylcarbamoyl, preferablyphenylcarbamoyl; (C₁-C₆)alkaneacylamino, preferably acetylamino;(C₆-C₁₀)areneacylamino, preferably benzoylamino; (C₁-C₆)alkanesulfonyl,preferably methanesulfonyl; (C₁-C₆)alkanesulfonamido, preferablymethanesulfonamido; (C₆-C₁₀)arenesulfonyl, preferably benzenesulfonyl ortoluenesulfonyl; (C₆-C₁₀)arenesulfonamido, preferably benzenesulfonyl ortoluenesulfonyl; (C₆-C₁₀)ar(C₁-C₆)alkylsulfonamido, preferablybenzylsulfonamido; C₁-C₆ alkylcarbonyl, preferably C₁-C₃ alkylcarbonyl,more preferably acetyl; (C₁-C₆)acyloxy, preferably acetoxy; cyano;amino; carboxy; hydroxy; ureido; and nitro. One or more carbon atoms ofan alkyl, cycloalkyl, or heterocyclyl group may also be optionallysubstituted with an oxo group.

In some particularly preferred embodiments, Cy is a phenyl, naphthyl,thienyl, benzothienyl, or quinolyl moiety which is unsubstituted or issubstituted by one or two substituents independently selected from thegroup consisting of C₁-C₄ alkyl, C₁-C₄ haloalkyl, C₆-C₁₀ aryl,(C₆-C₁₀)ar(C₁-C₆)alkyl, halo, nitro, hydroxy, C₁-C₆ alkoxy, C₁-C₆alkoxycarbonyl, carboxy, and amino

In some preferred embodiments, Z is −O—C(O)—R¹⁰,—O—C(O)-[C(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³) or —OR¹¹.

In some preferred embodiments, the amino acid is an L-amino acid.

In certain preferred embodiments, the sugar residue is a saccharideselected from the group consisting of glucose, galactose, mannose,gulose, idose, talose, allose, altrose, fructose, rhamnose, ribose andxylose.

In a second embodiment, the invention provides prodrugs of inhibitors ofhistone deacetylase, the prodrugs represented by formula (2):

Cy-L²-Ar-Y²—C(O)N(R^(x))—Z  (2)

and pharmaceutically acceptable salts thereof, wherein

Cy is H, cycloalkyl, aryl, heteroaryl, or heterocyclyl, any of which maybe optionally substituted, provided that Cy is not a(spirocycloalkyl)heterocyclyl;

L² is C₁-C₆ saturated alkylene or C₂-C₆ alkenylene, wherein the alkyleneor alkenylene optionally may be substituted, and wherein one or two ofthe carbon atoms of the alkylene is optionally replaced by aheteroatomic moiety independently selected from the group consisting of0; NR′, R′ being alkyl, acyl, or hydrogen; S; S(O); or S(O)₂;

Ar is arylene, wherein said arylene optionally may be additionallysubstituted and optionally may be fused to an aryl or heteroaryl ring,or to a saturated or partially unsaturated cycloalkyl or heterocyclicring, any of which may be optionally substituted; and

Y² is a chemical bond or a straight- or branched-chain saturatedalkylene, which may be optionally substituted, provided that thealkylene is not substituted with a substituent of the formula —C(O)Rwherein R comprises an α-amino acyl moiety;

R^(x) is H or —OH;

Z is —R²⁰, —O—R²⁰, —R²¹, or

wherein —R²⁰ is selected from the group consisting of —C(O)—R¹⁰,—C(O)O—R¹⁰, —R¹¹, —CH(R¹²)—O—C(O)—R¹⁰,—C(O)—C[(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³),—S(O₂)R¹⁰—C(O)—(CH₂)₁₋₄—C(OH)(COOR¹⁰)—(CH₂)₁₄—COOR¹⁰,—C(O)-[C(R¹⁴)(R¹⁴)]₁₋₄—P(O)(OH)(OH),—C(O)—(CH₂)₁₋₄—N(R¹⁴)—C[═N(R^(10′))]-N(R^(10′))(R^(10′)),—C(O)—(CH₂)—CH(OH)—(CH₂)—N(CH₃)(CH₃), —C(O)—CH(NH₂)—(CH₂)₁₋₆—COOH(preferably —C(O)—CH(NH₂)—(CH₂)—COOH), —C(O)—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰,—C(O)—O—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰ and —C(O)—(CH₂)_(n)—C(O)OR¹⁰,provided that the N to which Z is bound is not directly bonded to two Oatoms; and further provided that (a) when Z is —R²⁰ then R^(x) is —OH,and (b) when Z is —OR²⁰ then R^(x) is —H; or

R^(x) is absent and R²⁰ forms an optionally substituted heterocyclicring with the N to which it is attached;

n is 0, 1, 2, 3, or 4, preferably 1, 2, 3, or 4;

each R¹⁰ is independently selected from the group consisting ofhydrogen, optionally substituted C₁-C₂₀ alkyl, optionally substitutedC₂-C₂₀ alkenyl, optionally substituted C₂-C₂₀ alkynyl, optionallysubstituted C₁-C₂₀ alkoxycarbonyl, optionally substituted cycloalkyl,optionally substituted heterocycloalkyl, optionally substituted aryl,optionally substituted heteroaryl, optionally substitutedcycloalkylalkyl, optionally substituted heterocycloalkylalkyl,optionally substituted arylalkyl, optionally substitutedheteroarylalkyl, optionally substituted cycloalkylalkenyl, optionallysubstituted heterocycloalkylalkenyl, optionally substituted arylalkenyl,optionally substituted heteroarylalkenyl, optionally substitutedcycloalkylalkynyl, optionally substituted heterocycloalkylalkynyl,optionally substituted arylalkynyl, optionally substitutedheteroarylalkynyl, optionally substituted alkylcycloalkyl, optionallysubstituted alkylheterocycloalkyl, optionally substituted alkylaryl,optionally substituted alkylheteroaryl, optionally substitutedalkenylcycloalkyl, optionally substituted alkenylheterocycloalkyl,optionally substituted alkenylaryl, optionally substitutedalkenylheteroaryl, optionally substituted alkynylcycloalkyl, optionallysubstituted alkynylheterocycloalkyl, optionally substituted alkynylary,optionally substituted alkynylheteroaryl, a sugar residue, and an aminoacid residue (preferably bonded through the carboxy terminus of theamino acid);

each R^(10′) is independently hydrogen or C₁₋₆alkyl, or

R¹⁰ and R^(10′) together with the carbon atom to which they are attachedform an optionally substituted spirocycloalkyl;

R²¹ is a sugar or -amino acid-R¹³, wherein R¹³ is covalently bound tothe N-terminus;

R¹¹ is selected from the group consisting of hydrogen, optionallysubstituted heterocycloalkyl, optionally substituted aryl, andoptionally substituted heteroaryl;

R¹² is selected from hydrogen or alkyl; and

R¹³ is selected from the group consisting of hydrogen,—C(O)—CH[N(R^(10′))(R^(10′))]C₁-C₆alkyl,—C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-N(R^(10′))(R^(10′))C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-aryl,—C(O)—CH[N(R^(10′))(R^(10′))]—C₁-C₆alkyl-heteroaryl, —C(O)-aryl,—C(O)-heteroaryl, an amino protecting group, and R¹⁰; and

each R¹⁴ is independently selected from the group consisting of H,C₁-C₆alkyl and cycloalkyl, or two R¹⁴, together with the atom to whichthey are attached, form a cycloalkyl.

Preferred substituents Cy, Ar, and Z according to this aspect of theinvention are as defined above for the first embodiment. Preferredsubstituents of Y² are as defined above for Y¹. In some preferredembodiments, L² is saturated C₁-C₈ alkylene, more preferably C₁-C₆alkylene, still more preferably C₁-C₄ alkylene, any of which groups maybe optionally substituted. In some other preferred embodiments, L² isC₂-C₈ alkenylene, more preferably C₂-C₆ alkenylene, and still morepreferably C₂-C₄ alkenylene, any of which groups may be optionallysubstituted. The alkylene or alkenylene group may be substituted at oneor more carbon positions with a substituent preferably selected from thelist of preferred substituents recited above. More preferably, L² issubstituted at one or two positions with a substituent independentlyselected from the group consisting of C₁-C₆ alkyl, C₆-C₁₀ aryl, amino,oxo, hydroxy, C₁-C₄ alkoxy, and C₆-C₁₀ aryloxy. In some particularlypreferred embodiments, the alkylene or alkenylene group is substitutedwith one or two oxo or hydroxy groups.

In some preferred embodiments, L¹ is C₁-C₆ saturated alkylene, whereinon of the carbon atoms of the saturated alkylene is replaced by aheteroatom moiety selected from the group consisting of O; NR′, R′ beingalkyl, acyl, or hydrogen; S; S(O); or S(O)₂. Preferably, the carbon atomadjacent to Cy is replaced by a heteroatom moiety. In some particularlypreferred embodiments, L¹ is selected from the group consisting of—S—(CH₂)₂—; —S(O)—(CH₂)₂—, —S(O)₂—(CH₂)₂—, —S—(CH₂)₃—, —S(O)—(CH₂)₃—,and —S(O)₂—(CH₂)₃—.

In some preferred embodiments, Z is —O—C(O)—R¹⁰,—O—C(O)-[C(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³) or —OR¹¹. Even more preferredembodiments of compound (2) are:

In a third embodiment, the invention provides prodrugs of inhibitors ofhistone deacetylase, the prodrugs represented by formula (3):

Cy-L³-Ar-Y³—C(O)N(R^(x))—Z  (3)

and pharmaceutically acceptable salts thereof, wherein

Cy is —H, cycloalkyl, aryl, heteroaryl, or heterocyclyl, any of whichmay be optionally substituted, provided that Cy is not a(spirocycloalkyl)heterocyclyl;

L³ is selected from the group consisting of

(a) —(CH₂)_(m)—W—, where m is 0, 1, 2, 3, or 4, and W is selected fromthe group consisting of —C(O)NH—, —S(O)₂NH—, —NHC(O)—, —NHS(O)₂—, and—NH—C(O)—NH—; and

(b) C₁-C₆ alkylene or C₂-C₆ alkenylene, wherein the alkylene oralkenylene optionally may be substituted, and wherein one of the carbonatoms of the alkylene optionally may be replaced by O; NR′, R′ beingalkyl, acyl, or hydrogen; S; S(O); or S(O)₂;

Ar is arylene, wherein said arylene optionally may be additionallysubstituted and optionally may be fused to an aryl or heteroaryl ring,or to a saturated or partially unsaturated cycloalkyl or heterocyclicring, any of which may be optionally substituted; and

Y³ is C₂ alkenylene or C₂ alkynylene, wherein one or both carbon atomsof the alkenylene optionally may be substituted with alkyl, aryl,alkaryl, or aralkyl;

R^(x) is H or —OH;

Z is —R²⁰, —O—R²⁰, —R²¹, or

wherein —R²⁰ is selected from the group consisting of —C(O)—R¹⁰,—C(O)O—R¹⁰, —R¹¹, —CH(R¹²)—O—C(O)—R¹⁰,—C(O)—(CH₂)₁₋₄—C(OH)(COOR¹⁰)—(CH₂)₁₋₄—COOR¹⁰,—C(O)-[C(R¹⁴)(R¹⁴)]₁₋₄—P(O)(OH)(OH),—C(O)—(CH₂)₁₋₄—N(R¹⁴)—C[═N(R^(10′))]-N(R^(10′))(R^(10′))—C(O)—(CH₂)—CH(OH)—(CH₂)—N(CH₃)(CH₃),—C(O)—CH(NH₂)—(CH₂)₁₋₆—COOH (preferably —C(O)—CH(NH₂)—(CH₂)—COOH),—C(O)—C[(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³), —S(O₂)R¹⁰, —P(O)(OR¹⁰)(OR¹⁰),—C(O)—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰, —C(O)—O—(CH₂)_(n)—CH(OH)—CH₂—O—R¹⁰ and—C(O)—(CH₂)_(n)—C(O)OR¹⁰, provided that the N to which Z is bound is notdirectly bonded to two O atoms; and further provided that (a) when Z is—R²⁰ then R^(x) is —OH, and (b) when Z is —OR²⁰ then R^(x) is —H; or

R^(x) is absent and R²⁰ forms an optionally substituted heterocyclicring with the N to which it is attached;

n is 0, 1, 2, 3, or 4, preferably 1, 2, 3, or 4;

each R¹⁰ is independently selected from the group consisting ofhydrogen, optionally substituted C₁-C₂₀ alkyl, optionally substitutedC₂-C₂₀ alkenyl, optionally substituted C₂-C₂₀ alkynyl, optionallysubstituted C₁-C₂₀ alkoxycarbonyl, optionally substituted cycloalkyl,optionally substituted heterocycloalkyl, optionally substituted aryl,optionally substituted heteroaryl, optionally substitutedcycloalkylalkyl, optionally substituted heterocycloalkylalkyl,optionally substituted arylalkyl, optionally substitutedheteroarylalkyl, optionally substituted cycloalkylalkenyl, optionallysubstituted heterocycloalkylalkenyl, optionally substituted arylalkenyl,optionally substituted heteroarylalkenyl, optionally substitutedcycloalkylalkynyl, optionally substituted heterocycloalkylalkynyl,optionally substituted arylalkynl, optionally substitutedheteroarylalkynyl, optionally substituted alkylcycloalkyl, optionallysubstituted alkylheterocycloalkyl, optionally substituted alkylaryl,optionally substituted alkylheteroaryl, optionally substitutedalkenylcycloalkyl, optionally substituted alkenylheterocycloalkyl,optionally substituted alkenylaryl, optionally substitutedalkenylheteroaryl, optionally substituted alkynylcycloalkyl, optionallysubstituted alkynylheterocycloalkyl, optionally substituted alkynylary,optionally substituted alkynylheteroaryl, a sugar residue, and an aminoacid residue (preferably bonded through the carboxy terminus of theamino acid);

each R^(10′) is independently hydrogen or C₁₋₆alkyl, or

R¹⁰ and R^(10′) together with the carbon atom to which they are attachedform an optionally substituted spirocycloalkyl;

R²¹ is a sugar or -amino acid-R¹³, wherein R¹³ is covalently bound tothe N-terminus;

R¹¹ is selected from the group consisting of hydrogen, optionallysubstituted heterocycloalkyl, optionally substituted aryl, andoptionally substituted heteroaryl;

R¹² is selected from hydrogen or alkyl; and

R¹³ is selected from the group consisting of hydrogen,—C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl,—C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-N(R¹⁰)(R^(10′)),—C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-aryl,—C(O)—CH[N(R^(10′))(R^(10′))]-C₁-C₆alkyl-heteroaryl, —C(O)-aryl,—C(O)-heteroaryl, an amino protecting group, and R¹⁰; and

each R¹⁴ is independently selected from the group consisting of H,C₁-C₆alkyl and cycloalkyl, or two R¹⁴, together with the atom to whichthey are attached, form a cycloalkyl.

Preferred substituents Cy, Ar, and Z according to this aspect of theinvention are as defined above for the first embodiment. Preferredsubstituents L³ are as defined above for L¹ or L².

Preferably, Y³ is C₂ alkenylene or C₂ alkynylene, wherein one or bothcarbon atoms of the alkenylene optionally may be substituted with C₁-C₆alkyl, C₆-C₁₀ aryl, (C₁-C₆)alk(C₆-C₁₀)aryl, or (C₆-C₁₀)ar(C₁-C₆)alkyl.More preferably, Y³ is C₂ alkenylene or C₂ alkynylene, wherein one orboth carbon atoms of the alkenylene optionally may be substituted withC₁-C₄ alkyl, C₆-C₁₀ aryl, (C₁-C₄)alk(C₆-C₁₀)aryl, or(C₆-C₁₀)ar(C₁-C₄)alkyl. Still more preferably, Y³ is selected from thegroup consisting of —C≡C—, —CH═CH—, —C(CH₃)═CH—, and —CH═C(CH₃)—.

In a preferred embodiment of the compounds of formulae (1), (2), and(3), Z is selected from the group consisting of—O—C(O)—(CH₂)₁₋₄—C(OH)(COOR¹⁰)—(CH₂)₁₄—COOR¹⁰,—O—C(O)-[C(R¹⁴)(R¹⁴)]₁₋₄—P(O)(OH)(OH),—O—C(O)—(CH₂)₁₄—N(R¹⁴)—C[═N(R^(10′))]-N(R^(10′))(R^(10′)),—O—C(O)—(CH₂)—CH(OH)—(CH₂)—N(CH₃)(CH₃), —O—C(O)—CH(NH₂)—(CH₂)₁₋₆—COOH,preferably —O—C(O)—CH(NH₂)—(CH₂)—COOH.

In a preferred embodiment of the compounds of formulae (1), (2), and(3), Z is selected from the group consisting of—O—C(O)—(CH₂)—C(OH)(COOH)—(CH₂)—COOH, —O—C(O)—CH₂—P(O)(OH)(OH),—O—C(O)—(CH₂)—N(CH₃)—C(═NH)—NH₂, —O—C(O)—(CH₂)—CH(OH)—(CH₂)—N(CH₃)(CH₃),—O—C(O)—CH(NH₂)—(CH₂)—COOH.

In some preferred embodiments, Z is —O—C(O)—R¹⁰,—O—C—(O)-[C(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³) or —OR¹¹.

In some preferred embodiments of the prodrugs of inhibitors of histonedeacetylase, of formulae (1), (2), and (3), Z is —O—R²⁰ wherein R²⁰ is—C(O)—CR¹⁰R^(10′)—NH(R¹³), R¹³ and R^(10′) are H, and R¹⁰ is C₁-C₆-alkylor an amino acid side chain, or R¹⁰ and R^(10′) together with the carbonto which they are linked form C₃-C₆ cycloalkyl.

Naturally-occurring or non-naturally occurring amino acids are used toprepare the prodrugs of the invention. In particular, standard aminoacids suitable as a prodrug moiety include valine, leucine, isoleucine,methionine, phenylalanine, asparagine, glutamic acid, glutamine,histidine, lysine, arginine, aspartic acid, glycine, alanine, serine,threonine, tyrosine, tryptophan, cysteine and proline. Particularlypreferred are L-amino acids. Optionally an included amino acid is an α-,β-, or γ-amino acid. Also, naturally-occurring, non-standard amino acidscan be utilized in the compositions and methods of the invention. Forexample, in addition to the standard naturally occurring amino acidscommonly found in proteins, naturally occurring amino acids alsoillustratively include 4-hydroxyproline, gamma.-carboxyglutamic acid,selenocysteine, desmosine, 6-N-methyllysine,epsilon.-N,N,N-trimethyllysine, 3-methylhistidine, O-phosphoserine,5-hydroxylysine, epsilon.-N-acetyllysine, omega.-N-methylarginine,N-acetylserine, gamma-aminobutyric acid, citrulline, ornithine,azaserine, homocysteine, beta.-cyanoalanine and S-adenosylmethionine.Non-naturally occurring amino acids include phenyl glycine,meta-tyrosine, para-amino phenylalanine, 3-(3-pyridyl)-L-alanine-,4-(trifluoromethyl)-D-phenylalanine, and the like.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (1), (2) and (3) as definedabove, except that R²⁰ of Z is described in U.S. Pat. No. 4,443,435(incorporated by reference in its entirety) as comprising—CH(R¹³⁰)—X—C(O)—R¹³¹ wherein

X is O, S, or NR¹³²;

R¹³¹ is

-   -   (a) straight or branched chain alkyl having from 1 to 20 carbon        atoms especially methyl, ethyl, isopropyl, t-butyl, pentyl or        hexyl;    -   (b) aryl having from 6 to 10 carbon atoms especially phenyl,        substituted penyl or naphthalene;    -   (c) cycloalkyl having from 3 to 8 carbon atoms especially        cyclopentyl, or cyclohexyl;    -   (d) alkenyl having from 2-20 carbon atoms especially C₂₋₆        alkenyl such as vinyl, allyl, or butenyl;    -   (e) cycloalkenyl having from 5 to 8 carbon atoms especially        cyclopentenyl or cyclohexenyl;    -   (f) alkynyl having from 2 to 20 carbon atoms especially C₂₋₆        alkynyl for example, ethynyl, propynyl or hexynyl;    -   (g) aralkyl, alkaryl, aralkenyl, aralkynyl, alkenylaryl or        alkynylaryl wherein alkyl, aryl, alkenyl and alkynyl are as        previously defined;    -   (h) loweralkoxycarbonyl especially C₁₋₆ alkoxycarbonyl such as        methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl and        cyclopentoxycarbonyl;    -   (i) carboxyalkyl or alkanoyloxyalkyl especially carboxy-C₁₋₆        alkyl such as formyloxymethyl and formyloxypropyl; or C₁₋₆        (alkylcarboxyalkyl) such as acetoxymethyl, n-propanoyloxyethyl        and pentanoyloxybutyl;    -   (j) saturated or unsaturated monoheterocyclic or        polyheterocyclic, or fused heterocyclic, either directly bonded        to the carbonyl function or linked thereto via an alkylene        bridge, containing from 1 to 3 of any one or more of the        heteroatoms N, S or O in each heterocyclic ring thereof and each        such ring being from 3- to 8-membered; and    -   (k) mono- or polysubstituted derivatives of the above, each of        said substituents being selected from the group consisting of        lower alkyl; lower alkoxy; lower alkanoyl; lower alkanoyloxy;        halo especially bromo, chloro, or fluoro; haloloweralkyl        especially fluoro, chloro or bromoloweralkyl such as        trifluoromethyl and 1-chloropropyl; cyano; carbethoxy;        loweralkylthio, especially C₁₋₆ loweralkylthio such as        methylthio, ethylthio and n-propylthio; nitro; carboxyl; amino;        loweralkylamino especially C₁₋₆ alkylamino, for example,        methylamino, ethylamino and n-butylamino; diloweralkylamino        especially di(C₁₋₆ loweralkyl)amino such as N,N-dimethylamino,        N,N-diethylamino and N,N-dihexylamino; carbamyl;        loweralkylcarbamyl especially C₁₋₆ alkylcarbamyl such as        methylcarbamyl and ethyl carbamoyl; and R¹³³—X—C(O)-phenyl-,        wherein R¹³³ is hydrogen or alkyl having from 1 to 10 carbons;

R¹³⁰ is hydrogen, (b) R¹³¹, lower alkanoyl, cyano, haloloweralkyl,carbamyl, loweralkylcarbamyl, or diloweralkylcarbamyl, —CH₂ONO₂, or—CH₂OCOR¹³¹;

R¹³² is hydrogen or lower alkyl; and further wherein R¹³¹ and R¹³⁰ maybe taken together to form a ring cyclizing moiety selected from thegroup consisting of:

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (1), (2) and (3) as definedabove, except that R²⁰ of Z is described in U.S. Pat. No. 6,407,235(incorporated by reference in its entirety) as comprising:

a) —C(O)(CH₂)_(m)C(O)OR⁴⁰, wherein m is 1, 2, 3 or 4,

-   -   b)

wherein R⁴¹ is —N(R⁴²)(R⁴³) and R⁴² and R⁴³ are hydrogen or lower alkyl,or is a five or six member heterocyclyl or heteroaryl optionallysubstituted by lower alkyl, or

c) —C(O)(CH₂)NHC(O)(CH₂)N(R⁴²)(R⁴³).

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (1), (2) and (3) as definedabove, except that R²⁰ of Z is described in U.S. Pat. No. 6,545,131(incorporated by reference in its entirety) as comprising:CO—(CH═CH)_(n1)—(CH₂)_(n2)—Ar—NH₂, —CO—(CH₂)_(n2)—(CH═CH)_(n1)—Ar—NH₂,CO—(CH₂)_(n2)—(CH═CH)_(n1)—CO—NH—Ar—NH₂ andCO—(CH═CH)_(n1)—(CH₂)_(n2)—CO—NH—Ar—NH₂ and substituted variationsthereof, where n1 and n2 are from 0 to 5, Ar is a substituted orunsubstituted aryl group. In some preferred embodiments, Z isCO—(CH₂)_(n3)—NH₂, where n3 is from 0 to 15, preferably 3-15, and alsopreferably 6-12. Particularly preferred substituent groups within thisclass are 6-aminohexanoyl, 7-aminoheptanoyl, 8-aminooctanoyl,9-aminononanoyl, 10-aminodecanoyl, 11-aminoundecanoyl, and12-aminododecanoyl. These substituents are generally synthesized fromthe corresponding amino acids, 6-aminohexanoic acid, and so forth. Theamino acids are N-terminal protected by standard methods, for exampleBoc protection. Dicyclohexylcarbodiimide (DCCI)-promoted coupling of theN-terminal protected substituent to thapsigargin, followed by standarddeprotection reactions produces primary amine-containing thapsigarginanalogs.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (1), (2) and (3) as definedabove, except that R²⁰ of Z is described in U.S. Pat. No. 7,115,573(incorporated by reference in its entirety) as comprising:

-   -   (1) an oligopeptide of the formula (AA)_(n)-AA³-AA²-AA¹,        wherein: each AA independently represents an amino acid, n is 0        or 1, and when n is 1, then (AA)_(n) is AA⁴ which represents any        amino acid, AA³ represents isoleucine, AA² represents any amino        acid, and AA¹ represents any amino acid,    -   (2) a stabilizing group, and    -   (3) optionally, a linker group not cleavable by a trouase, such        as TOP (described in greater detail below)

wherein the oligopeptide is directly linked to the stabilizing group ata first attachment site of the oligopeptide and the oligopeptide isdirectly linked to the therapeutic agent or indirectly linked throughthe linker group to the therapeutic agent at a second attachment site ofthe oligopeptide,

wherein the stabilizing group hinders cleavage of the compound byenzymes present in whole blood, and

wherein the compound is cleavable by an enzyme associated with thetarget cell, the enzyme associated with the target cell being other thanTOP (Thimet oligopeptidase). The compound preferably includes anoligopeptide that is resistant to cleavage by a trouase, particularlyTOP, i.e., resistant to cleavage under physiological conditions. Theoptionally present linker group that is not cleavable by a trouase isnot cleavable under physiological conditions.

The typical orientation of these portions of the prodrug is as follows:(stabilizing group)-(oligopeptide)-(optional linker group)-(therapeuticagent).

Direct linkage of two portions of the prodrug means a covalent bondexists between the two portions. The stabilizing group and theoligopeptide are therefore directly linked via a covalent chemical bondat the first attachment site of the oligopeptide, typically theN-terminus of the oligopeptide. When the oligopeptide and thetherapeutic agent are directly linked then they are covalently bound toone another at the second attachment site of the oligopeptide. Thesecond attachment site of the oligopeptide is typically the C-terminusof the oligopeptide, but may be elsewhere on the oligopeptide.

Indirect linkage of two portions of the prodrug means each of the twoportions is covalently bound to a linker group. In an alternativeembodiment, the prodrug has indirect linkage of the oligopeptide to thetherapeutic agent. Thus, typically, the oligopeptide is covalently boundto the linker group which, in turn, is covalently bound to thetherapeutic agent.

In an alternative embodiment, the orientation of the prodrug may bereversed so that a stabilizing group is attached to the oligopeptide atthe C-terminus and the therapeutic agent is directly or indirectlylinked to the N-terminus of the oligopeptide. Thus, in an alternativeembodiment, the first attachment site of the oligopeptide may be theC-terminus of the oligopeptide and the second attachment site by theoligopeptide may be the N-terminus of the oligopeptide. The linker groupmay optimally be present between the therapeutic agent and theoligopeptide. The alternative embodiment of the prodrug of the inventionfunctions in the same manner as does the primary embodiment.

The stabilizing group typically protects the prodrug from cleavage byproteinases and peptidases present in blood, blood serum, and normaltissue. Particularly, since the stabilizing group caps the N-terminus ofthe oligopeptide, and is therefore sometimes referred to as an N-cap orN-block, it serves to ward against peptidases to which the prodrug mayotherwise be susceptible. A stabilizing group that hinders cleavage ofthe oligopeptide by enzymes present in whole blood is chosen from thefollowing:

-   -   (1) other than an amino acid, or    -   (2) an amino acid that is either (i) a non-genetically-encoded        amino acid or (ii) aspartic acid or glutamic acid attached to        the N-terminus of the oligopeptide at the β-carboxyl group of        aspartic acid or the γ-carboxyl group of glutamic acid.

For example, dicarboxylic (or a higher order carboxylic) acid or apharmaceutically acceptable salt thereof may be used as a stabilizinggroup. Since chemical radicals having more than two carboxylic acids arealso acceptable as part of the prodrug, the end group havingdicarboxylic (or higher order carboxylic) acids is an exemplary N-cap.The N-cap may thus be a monoamide derivative of a chemical radicalcontaining two or more carboxylic acids where the amide is attached ontothe amino terminus of the peptide and the remaining carboxylic acids arefree and uncoupled. For this purpose, the N-cap is preferably succinicacid, adipic acid, glutaric acid, or phthalic acid, with succinic acidand adipic acid being most preferred. Other examples of useful N-caps inthe prodrug compound of the invention include diglycolic acid, fumaricacid, naphthalene dicarboxylic acid, pyroglutamic acid, acetic acid, 1-or 2-, naphthylcarboxylic acid, 1,8-naphthyl dicarboxylic acid, aconiticacid, carboxycinnamic acid, triazole dicarboxylic acid, gluconic acid,4-carboxyphenyl boronic acid, a (PEG)_(n)-analog such as polyethyleneglycolic acid, butane disulfonic acid, maleic acid, nipecotic acid, andisonipecotic acid.

Further, a non-genetically encoded amino acid such as one of thefollowing may also be used as the stabilizing group: β-Alanine,Thiazolidine-4-carboxylic acid, 2-Thienylalanine, 2-Naphthylalanine,D-Alanine, D-Leucine, D-Methionine, D-Phenylalanine,3-Amino-3-phenylpropionic acid, γ-Aminobutyric acid,3-amino-4,4-diphenylbutyric acid, Tetrahydroisoquinoline-3-carboxylicacid, 4-Aminomethylbenzoic acid, and Aminoisobutyric acid.

A linker group between the oligopeptide and the therapeutic agent may beadvantageous for reasons such as the following: 1. As a spacer forsteric considerations in order to facilitate enzymatic release of theAA¹ amino acid or other enzymatic activation steps. 2. To provide anappropriate attachment chemistry between the therapeutic agent and theoligopeptide. 3. To improve the synthetic process of making the prodrugconjugate (e.g., by pre-derivitizing the therapeutic agent oroligopeptide with the linker group before conjugation to enhance yieldor specificity.) 4. To improve physical properties of the prodrug. 5. Toprovide an additional mechanism for intracellular release of the drug.

Linker structures are dictated by the required functionality. Examplesof potential linker chemistries are hydrazide, ester, ether, andsulfhydryl Amino caproic acid is an example of a bifunctional linkergroup. When amino caproic acid is used as part of the linker group, itis not counted as an amino acid in the numbering scheme of theoligopeptide.

The oligopeptide moiety is linked at a first attachment site of theoligopeptide to a stabilizing group that hinders cleavage of theoligopeptide by enzymes present in whole blood, and directly orindirectly linked to a therapeutic agent at a second attachment site ofthe oligopeptide. The linkage of the oligopeptide to the therapeuticagent and the stabilizing group may be performed in any order orconcurrently. The resulting conjugate is tested for cleavability by TOP.Test compounds resistant to cleavage by TOP are selected. The resultingconjugate may also be tested for stability in whole blood. Testcompounds stable in whole blood are selected.

The combination of oligopeptide, stabilizing group, and optional linkerof U.S. Pat. No. 7,115,573 is further described in US 2002-0142955, alsoincorporated herein by reference.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (1), (2) and (3) as definedabove, except that R²⁰ of Z is described in US 2004-0019017 A1(incorporated by reference in its entirety and which describes caspaseinhibitor prodrugs), as comprising:

wherein R⁵¹ is a saturated or unsaturated, straight-chain or branched,substituted or unsubstituted alkyl of 2 to 30, preferably 2 to 24,carbon atoms;

R⁵² is H or a phospholipid head group, preferably choline;

X is a direct covalent bond or a group C(O)LR⁵³ wherein L is a saturatedor unsaturated, straight-chain or branched, substituted or unsubstitutedalkyl having from 2 to 15 carbon atoms, which optionally includes cyclicelements, and is optionally interrupted by one or more atoms selectedfrom the group consisting of oxygen, sulfur and N(R⁵⁴); R⁵³ is selectedfrom the group consisting of O, S and N(R⁵⁴), wherein R⁵⁴ is H or asaturated or unsaturated alkyl having 1 to 6 carbon atoms.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (1), (2) and (3) as definedabove, except that R²⁰ of Z is the Y moiety described in U.S. Pat. No.7,115,573 (incorporated by reference in its entirety).

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (1), (2) and (3) as definedabove, except that R²⁰ of Z is described in US 2006-0166903 A1(incorporated by reference in its entirety, ascomprising-X-L-O—P(O)(O⁻)—O—CH₂—CH₂—N(CH₃)₃ ⁺, wherein X and L are asdescribed in US 2006-0166903A1.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (1), (2) and (3) as definedabove, except Z is one of the cleavable prodrug moieties described inU.S. Pat. No. 6,855,702, US 2005-0137141, and US 2006-0135594, allhereby incorporated by reference in their entirety.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (1), (2) and (3) as definedabove, wherein

Cy is optionally substituted aryl, preferably optionally substitutedphenyl;

Ar is optionally substituted aryl, preferably optionally substitutedphenyl;

R^(x) is H or OH; and

Z is —O—R²⁰ or R²¹.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (1), (2) and (3) as definedabove, wherein

Cy is optionally substituted aryl, preferably optionally substitutedphenyl;

Ar is optionally substituted aryl, preferably optionally substitutedphenyl;

Rx is H or OH; and

Z is —O—R²⁰ or R²¹, wherein

R²⁰ is —C(O)—C[(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³) or —C(O)—R¹⁰.

In a preferred embodiment of the present invention, the prodrugs ofinhibitors of histone deacetylase comprise those of formula (2).

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (2) as defined above,wherein

Cy is optionally substituted aryl, preferably optionally substitutedphenyl;

Ar is optionally substituted aryl, preferably optionally substitutedphenyl;

Rx is H or OH; and

Z is —O—R²⁰ or R²¹.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (2) as defined above,wherein

Cy is optionally substituted aryl, preferably optionally substitutedphenyl;

Ar is optionally substituted aryl, preferably optionally substitutedphenyl;

Rx is H or OH; and

Z is —O—R²⁰ or R²¹, wherein

R²⁰ is —C(O)—C[(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³) or —C(O)—R¹⁰.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (2) as defined above,wherein

-   -   Cy is optionally substituted aryl, preferably optionally        substituted phenyl, wherein the substituents are preferably        selected from the group consisting of —CF₃, halo, heterocyclyl        and fused heterocyclyl;    -   L² is saturated C₃alkyl or C₄alkyl, preferably unsubstituted;    -   Ar is optionally substituted aryl, preferably optionally        substituted phenyl;    -   Y² is C₁alkyl or C₂alkyl, preferably C₁alkyl, optionally        substituted;    -   Rx is H or OH, preferably H;    -   Z is —O—R²⁰ or R²¹;    -   R²⁰ is —C(O)—C[(R¹⁰)(R^(10′))]₁₋₄—NH(R) or —C(O)—R¹⁰;    -   each R¹⁰ is independently selected from the group consisting of        H, optionally substituted alkyl, optionally substituted        -alkylphenyl, optionally substituted -alkylheteroaryl and        optionally substituted heteroaryl;    -   each R^(10′) is independently H or alkyl; or    -   R¹⁰ and R^(10′) together with the atom to which they are        attached form a C₃ or C₄spirocycloalkyl, preferably a        C₃spirocycloalkyl;    -   R¹³ is selected from the group consisting of H,        —C(O)—CH[N(R¹⁰)(R^(10′))]-C₁-C₆alkyl-N(R¹⁰)(R^(10′)),        —C(O)-heteroaryl, —C(O)-aryl,        —C(O)—CH[N(R¹⁰)(R^(10′))]-C₁-C₆alkyl,        —C(O)—CH[N(R¹⁰)(R^(10′))]-C₁-C₆alkyl-aryl and        —C(O)—CH[N(R¹⁰)(R^(10′))]-C₁-C₆alkyl-heteroaryl; and    -   R²¹ is amino acid-R¹³ (preferably the amino acid is lysine or        arginine).

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (2) as defined above,wherein

-   -   Cy is optionally substituted aryl, preferably optionally        substituted phenyl, wherein the substituents are preferably        selected from the group consisting of —CF₃, halo, heterocyclyl        and fused heterocyclyl;    -   L² is saturated C₃alkyl or C₄alkyl, preferably unsubstituted;    -   Ar is optionally substituted aryl, preferably optionally        substituted phenyl;    -   Y² is C₁alkyl or C₂alkyl, preferably C₁alkyl, optionally        substituted;    -   Rx is H or OH, preferably H;    -   Z is —O—R²⁰;    -   R²⁰ is —C(O)—C[(R¹⁰)(R^(10′))]₁₋₄—NH(R¹³), preferably        —C(O)—C[(R¹⁰)(R^(10′))]₁₋₂—NH(R¹³), more preferably        —C(O)—C[(R¹⁰)(R^(10′))]—NH(R¹³);    -   each R¹⁰ is independently selected from the group consisting of        H, optionally substituted alkyl and optionally substituted        -alkylphenyl;    -   each R^(10′) is independently H or alkyl; or    -   R¹⁰ and R^(10′) together with the atom to which they are        attached form a C₃ or C₄spirocycloalkyl, preferably a        C₃spirocycloalkyl; and    -   R¹³ is H.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (2) as defined above,wherein

-   -   Cy is optionally substituted aryl, preferably optionally        substituted phenyl, wherein the substituents are preferably        selected from the group consisting of —CF₃, halo, heterocyclyl        and fused heterocyclyl;    -   L² is saturated C₃alkyl or C₄alkyl, preferably unsubstituted;    -   Ar is optionally substituted aryl, preferably optionally        substituted phenyl;    -   Y² is C₁alkyl or C₂alkyl, preferably C₁alkyl, optionally        substituted;    -   Rx is H or OH, preferably H;    -   Z is R²¹;    -   R²¹ is amino acid-R¹³ (preferably the amino acid is lysine or        arginine); and    -   R¹³ is H.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (2) as defined above,wherein

-   -   Cy is optionally substituted aryl, preferably optionally        substituted phenyl, wherein the substituents are preferably        selected from the group consisting of —CF₃, halo, heterocyclyl        and fused heterocyclyl;    -   L² is saturated C₃alkyl or C₄alkyl, preferably unsubstituted;    -   Ar is optionally substituted aryl, preferably optionally        substituted phenyl;    -   Y² is C₁alkyl or C₂alkyl, preferably C₁alkyl, optionally        substituted;    -   Rx is H or OH, preferably H;    -   Z is —O—R²⁰;    -   R²⁰ is —C(O)—R¹⁰; and    -   R¹⁰ is selected from the group consisting of optionally        substituted alkyl, optionally substituted -alkylphenyl,        optionally substituted -alkylheteroaryl and optionally        substituted heteroaryl.

In other embodiments, the prodrugs of inhibitors of histone deacetylaseof the invention comprise those of formulae (2) as defined above,wherein

-   -   Cy is optionally substituted aryl, preferably optionally        substituted phenyl, wherein the substituents are preferably        selected from the group consisting of —CF₃, halo, heterocyclyl        and fused heterocyclyl;    -   L² is saturated C₃alkyl or C₄alkyl, preferably unsubstituted;    -   Ar is optionally substituted aryl, preferably optionally        substituted phenyl;    -   Y² is C₁alkyl or C₂alkyl, preferably C₁alkyl, optionally        substituted;    -   Rx is H or OH, preferably H;    -   Z is —O—R²⁰;    -   R²⁰ is —C(O)—C[(R¹⁰)(R^(10′))]-NH(R¹³), wherein R¹⁰ and R^(10′)        together with the atom to which they are attached form a C₃ or        C₄spirocycloalkyl, preferably a C₃spirocycloalkyl; and    -   R¹³ is selected from the group consisting of H,        —C(O)—CH[N(R¹⁰)(R^(10′))]-C₁-C₆alkyl-N(R¹⁰)(R^(10′)),        —C(O)-heteroaryl, —C(O)-aryl,        —C(O)—CH[N(R¹⁰)(R^(10′))]-C₁-C₆alkyl,        —C(O)—CH[N(R¹⁰)(R^(10′))]-C₁-C₆alkyl-aryl and        —C(O)—CH[N(R¹⁰)(R^(10′))]-C₁-C₆alkyl-heteroaryl, wherein R¹⁰ and        R^(10′) are each independently selected from H and C₁-C₆alkyl,        preferably H.

Preferred prodrugs of the invention include those in Table A:

TABLE A

Preferred prodrug compounds of the invention are cleavable (e.g.,hydrolysable) in mammalian and/or fungal pathogen cells into compounds(cleavage products) in which Z in formulae (1), (2), and (3) is —OH.Such cleavage products are active histone deacetylase inhibitors. Thus,according to another aspect, the invention provides compounds offormulae (1), (2), and (3) as defined above (and pharmaceuticallyacceptable salts thereof) with the exception that Z is —OH. Among thepreferred cleavage compounds are those with structure:

Preferred cleavage products of the prodrug compounds of the inventioninclude those in Table A in which Z is —OH.

All compounds of the invention, whether prodrug or correspondingcleavage product, can be racemic or diastereomerically orenantiomerically enriched. In addition, compounds of the invention,whether prodrug or corresponding cleavage product, can be in the form ofa hydrate, solvate, pharmaceutically acceptable salt, and/or complex.

Synthesis

Compounds of formula Cy-L¹-Ar-Y¹—C(O)—NH—O—H, wherein L¹ is —S(O)₂NH—,preferably may be prepared according to the synthetic routes depicted inSchemes 1-5. Accordingly, in certain preferred embodiments, compounds Iare preferably prepared according to the general synthetic routedepicted in Scheme 1. Thus, a sulfonyl chloride (II) is treated with anamine (III) in a solvent such as methylene chloride in the presence ofan organic base such as triethylamine Treatment of the crude productwith a base such as sodium methoxide in an alcoholic solvent such asmethanol effects cleavage of any dialkylated material and affords thesulfonamide (IV). Hydrolysis of the ester function in IV can be effectedby treatment with a hydroxide base, such as lithium hydroxide, in asolvent mixture such as tetrahydrofuran and methanol to afford thecorresponding acid (V).

In some embodiments, conversion of the acid V to the hydroxamic acid Imay be accomplished by coupling V with a protected hydroxylamine, suchas tetrahydropyranylhydroxylamine (NH₂OTHP), to afford the protectedhydroxamate VI, followed by acidic hydrolysis of VI to provide thehydroxamic acid I. The coupling reaction is preferably accomplished withthe coupling reagent dicyclohexylcarbodiimide (DCC) in a solvent such asmethylene chloride (Method A) or with the coupling reagent1-(3-dimethylaminopropyl)-3-ethylcarbodiimide in presence of N-hydroxybenzotriazole in an aprotic solvent such as dimethylformamide (MethodD). Other coupling reagents are known in the art and may also be usedfor this reaction. Hydrolysis of VI is preferably effected by treatmentwith an organic acid such as camphorsulfonic acid in a protic solventsuch as methanol.

Alternatively, in some other embodiments, acid V is converted to thecorresponding acid chloride, preferably by treatment with oxalicchloride, followed by the addition of a protected hydroxylamine such asO-trimethylsilylhydroxylamine in a solvent such as methylene chloride,which then provides the hydroxylamine I upon workup (Method C).

In still other embodiments, the ester IV is preferably treated withhydroxylamine in a solvent such as methanol in the presence of a basesuch as sodium methoxide to furnish the hydroxylamine I directly (MethodB).

Compounds of formula X and XIV preferably are prepared according to thegeneral procedure outlined in Scheme 2. Thus, an aminoaryl halide (VII)is treated with a sulfonyl chloride in presence of a base such astriethylamine, followed by treatment with an alkoxide base, to furnishthe sulfonamide VIII. One of skill in the art will recognize thatreverse sulfonamide analogs can be readily prepared by an analogousprocedure, treating a haloarenesulfonyl halide with an arylamine

Compound VIII is coupled with a terminal acetylene or olefinic compoundin the presence of a palladium catalyst such astetrakis(triphenylphosphine)palladium(0) in a solvent such aspyrrolidine to afford IX.

Oxidation of the compound of formula IX (X=CH₂OH), followed byhomologation of the resulting aldehyde using a Wittig type reagent suchas carbethoxymethylenetriphenylphosphorane in a solvent such asacetonitrile, gives the compound of formula XI. Basic hydrolysis of XI,such as by treatment with lithium hydroxide in a mixture of THF andwater, provides the acid XII. Hydrogenation of XII may preferably beperformed over a palladium catalyst such as Pd/C in a protic solventsuch as methanol to afford the saturated acid XIII Coupling of the acidXIII with an O-protected hydroxylamine such asO-tetrahydropyranylhydroxylamine is effected by treatment with acoupling reagent such as 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidein the presence of N-hydroxybenzotriazole (HOBT), orN,N-dicyclohexylcarbodiimide (DCC), in a solvent such as DMF, followedby deprotection to furnish the compound of general formula XIV.

The acid IX, wherein X=COOH, may be coupled directly with an O-protectedhydroxylamine such as O-tetrahydropyranylhydroxylamine, followed bydeprotection of the hydroxy protecting group to furnish the hydroxamicacid X.

Compounds of formula Cy-L¹-Ar—Y¹—C(O)—NH—O—H, wherein L¹ is —C(O)NH—,preferably may be prepared according to the synthetic routes analogousto those depicted in Schemes 1-2, substituting acid chloride startingmaterials for the sulfonyl chloride starting materials in those Schemes.

Compounds of the formula Cy-L²-Ar-Y²—C(O)—NH—O—H are preferably preparedaccording to the synthetic routes outlined in Schemes 3-5. Accordingly,in certain preferred embodiments, compounds of formulae XIX and XXI(L²=—C(O)—CH═CH— or —C(O)—CH₂CH₂—) preferably are prepared according tothe route described in Scheme 3. Thus, a substituted aryl acetophenone(XV) is treated with an aryl aldehyde (XVI) in a protic solvent such asmethanol in the presence of a base such as sodium methoxide to affordthe enone XVII.

The acid substituent of XVII (R=H) is coupled with an O-protectedhydroxylamine such as O-tetrahydropyranylhydroxylamine(R₁=tetrahydropyranyl) to afford the O-protected-N-hydroxybenzamideXVIII. The coupling reaction is preferably performed by treating theacid and hydroxylamine with dicyclohexylcarbodiimide in a solvent suchas methylene chloride or with1-(3-dimethylaminopropyl)-3-ethylcarbodiimide in the presence ofN-hydroxybenzotriazole in a solvent such as dimethylformamide. Othercoupling reagents are known in the art and may also be used in thisreaction. O-Deprotection is accomplished by treatment of XVIII with anacid such as camphorsulfonic acid in a solvent such as methanol toafford the hydroxamic acid XIX (L²=—C(O)—CH═CH—).

Saturated compounds of formula XXI (L²=—C(O)—CH₂CH₂—) are preferablyprepared by hydrogenation of XVII (R=Me) over a palladium catalyst, suchas 10% Pd/C, in a solvent such as methanol-tetrahydrofuran. Basichydrolysis of the resultant product XIX with lithium hydroxide, followedby N-hydroxy amide formation and acid hydrolysis as described above,then affords the hydroxamic acid XXI.

Compounds of formula XXVI (L²=—(CH₂)_(o+2)—) are preferably prepared bythe general procedures described in Schemes 4 and 5. Thus, in someembodiments, a terminal olefin (XXII) is coupled with an aryl halide(XXIII) in the presence of a catalytic amount of a palladium source,such as palladium acetate or tris(dibenzylideneacetone)dipalladium(0), aphosphine, such as triphenylphosphine, and a base, such astriethylamine, in a solvent such as acetonitrile to afford the coupledproduct XXIV. Hydrogenation, followed by N-hydroxyamide formation andacid hydrolysis, as described above, affords the hydroxamic acid XXVI.

Alternatively, in some other embodiments, a phosphonium salt of formulaXXVII is treated with an aryl aldehyde of formula XXVIII in the presenceof base, such as lithium hexamethyldisilazide, in a solvent, such astetrahydrofuran, to produce the compound XXIV. Hydrogenation, followedby N-hydroxyamide formation and acidic hydrolysis, then affords thecompounds XXVI.

Compounds of formula Cy-L-Ar-Y—C(O)—NH—Z, wherein L is L¹ or L², Y is Y¹or Y², and Z is anilinyl or pyridyl, are preferably prepared accordingto synthetic routes outlined in Scheme 6. An acid of formulaCy-L-Ar-Y—C(O)—OH (XXIX), prepared by one of the methods shown inSchemes 1-5, is converted to the corresponding acid chloride XXXaccording to standard methods, e.g., by treatment with sodium hydrideand oxalyl chloride. Treatment of XXX with 2-aminopyridine and atertiary base such as N-methylmorpholine, preferably in dichloromethaneat reduced temperature, then affords the pyridyl amide XXXI. In asimilar fashion, the acid chloride XXX may be treated with1,2-phenylenediamine to afford the anilinyl amide XXXII. Alternatively,the acid chloride XXX may be treated with a mono-protected1,2-phenylenediamine, such as 2-(t-BOC-amino)aniline, followed bydeprotection, to afford XXXII.

In another alternative procedure, the acid XXIX may be activated bytreatment with carbonyldiimidazole (CDI), followed by treatment with1,2-phenylenediamine and trifluoroacetic acid to afford the anilinylamide XXXII.

Compounds of formula XXXVIII (L²=—C(O)-alkylene-) preferably areprepared according to the general procedure depicted in Scheme 7. Thus,Aldol condensation of ketone XXXIII (R₁=H or alkyl) with aldehyde XXXIVaffords the adduct XXXV. The adduct XXXV may be directly converted tothe corresponding hydroxamic acid XXXVI, or may first undergohydrogenation to afford the saturated compound XXVII and then beconverted to the hydroxamic acid XXXVIII.

Compounds of formula Cy-L²-Ar-Y²—C(O)—NH—O—H, wherein one of the carbonatoms in L² is replaced with S, S(O), or S(O)₂ preferably are preparedaccording to the general procedure outlined in Scheme 8. Thus, thiolXXXIX is added to olefin XL to produce XLI. The reaction is preferablyconducted in the presence of a radical initiator such as2,2′-azobisisobutyronitrile (AIBN) or1,1′-azobis(cyclohexanecarbonitrile) (VAZO™). Sulfide oxidation,preferably by treatment with m-chloroperbenzoic acid (mCPBA), affordsthe corresponding sulfone, which is conveniently isolated afterconversion to the methyl ester by treatment with diazomethane. Esterhydrolysis then affords the acid XLII, which is converted to thehydroxamic acid XLIII according to any of the procedures describedabove. The sulfide XLI also may be converted directly to thecorresponding hydroxamic acid XLIV, which then may be selectivelyoxidized to the sulfoxide XLV, for example, by treatment with hydrogenperoxide and tellurium dioxide.

Alternatively, compounds of Cy-L²-Ar-Y²—C(O)—NH—O—H can be preparedaccording to Scheme 9. In Scheme 9, haloaryl acetic acid XLVI isesterified, by, for example, treatment with HCl in dioxane in thepresence of an alcohol such as methanol, to afford acetate XLVII.Paladium coupling of acetate XLVII with alkyne XLVIII with, for example(Ph₃P)₄Pd in DME and diethylamine in the presence of Cut, producesXLVIX, which is subsequently reduced under H₂ and, for example, Pd/C inmethanol, to afford XLVX. N-hydroxyamide formation and acid hydrolysis,as described above, then leads to XLVXI.

Compounds of formulas (1)-(3) can be prepared as depicted in Scheme 10.XLVXI is treated with an amino acid XLVXII under standard peptidecoupling conditions, such as, for example, EDC and HOBt in DMF, toafford the protected acetamide XLVXIII, which is subsequentlydeprotected to yield the prodrug XLVXIV.

Other compounds of formula (1)-(3) can be prepared by methods known bythose skilled in the art. Examples of such methods can be found in U.S.Pat. Nos. 4,443,435; 6,407,235; 6,545,131; 6,855,702; 7,115,573; UnitedStates Patent Application Nos. US 2002-0142955, US 2004-0019017, US2005-0137141, US 2006-0135594, US 2006-0166903 and internationalpublication WO 2005/097747, all of which are incorporated herein byreference.

Pharmaceutical Compositions

In a second aspect, the invention provides pharmaceutical compositionscomprising a prodrug of an inhibitor of histone deacetylase representedby any one of formulae (1)-(3) and a pharmaceutically acceptablecarrier, excipient, or diluent. Compounds of the invention (whether aprodrug or a hydrolzyation product) or compositions thereof may beformulated by any method well known in the art and may be prepared foradministration by any route, including, without limitation, parenteral,oral, sublingual, transdermal, topical, intranasal, intratracheal, orintrarectal. In certain preferred embodiments, compounds of theinvention (whether a prodrug or a hydrolzyation product) or compositionsthereof are administered intravenously in a hospital setting. In certainother preferred embodiments, administration may preferably be by theoral route.

The characteristics of the carrier will depend on the route ofadministration. As used herein, the term “pharmaceutically acceptable”means a non-toxic material that is compatible with a biological systemsuch as a cell, cell culture, tissue, or organism, and that does notinterfere with the effectiveness of the biological activity of theactive ingredient(s). Thus, compositions according to the invention maycontain, in addition to the inhibitor, diluents, fillers, salts,buffers, stabilizers, solubilizers, and other materials well known inthe art. The preparation of pharmaceutically acceptable formulations isdescribed in, e.g., Remington's Pharmaceutical Sciences, 18th Edition,ed. A. Gennaro, Mack Publishing Co., Easton, Pa., 1990.

Inhibition of Histone Deacetylase

In a third aspect, the invention provides a method of inhibiting histonedeacetylase in a cell, comprising contacting a cell in which inhibitionof histone deacetylase is desired with a prodrug of an inhibitor ofhistone deacetylase according to any of formulas (1)-(3).

Measurement of the enzymatic activity of a histone deacetylase can beachieved using known methodologies. For example, Yoshida et al., J.Biol. Chem., 265: 17174-17179 (1990), describes the assessment ofhistone deacetylase enzymatic activity by the detection of acetylatedhistones in trichostatin A treated cells. Taunton et al., Science, 272:408-411 (1996), similarly describes methods to measure histonedeacetylase enzymatic activity using endogenous and recombinant HDAC-1.Both of these references are hereby incorporated by reference in theirentirety.

In some preferred embodiments, the histone deacetylase inhibitorinteracts with and reduces the activity of all histone deacetylases inthe cell. In some other preferred embodiments according to this aspectof the invention, the histone deacetylase inhibitor interacts with andreduces the activity of fewer than all histone deacetylases in the cell.In certain preferred embodiments, the inhibitor interacts with andreduces the activity of one histone deacetylase (e.g., HDAC-1), but doesnot interact with or reduce the activities of other histone deacetylases(e.g., HDAC-2, HDAC-3, HDAC-4, HDAC-5, HDAC-6, HDAC-7, HDAC-8, HDAC-9,HDAC-10, and HDAC-11). As discussed below, certain particularlypreferred histone deacetylase inhibitors are those that interact withand reduce the enzymatic activity of a histone deacetylase that isinvolved in tumorigenesis. Certain other preferred histone deacetylaseinhibitors interact with and reduce the enzymatic activity of a fungalhistone deacetylase.

Preferably, the method according to the third aspect of the inventioncauses an inhibition of cell proliferation of the contacted cells. Thephrase “inhibiting cell proliferation” is used to denote an ability ofan inhibitor of histone deacetylase to retard the growth of cellscontacted with the inhibitor as compared to cells not contacted. Anassessment of cell proliferation can be made by counting contacted andnon-contacted cells using a Coulter Cell Counter (Coulter, Miami, Fla.)or a hemacytometer, or other appropriate method (which may depend on thecell type being counted) known to those of skill in the art. Where thecells are in a solid growth (e.g., a solid tumor or organ), such anassessment of cell proliferation can be made by measuring the growthwith calipers and comparing the size of the growth of contacted cellswith non-contacted cells.

Preferably, growth of cells contacted with the prodrug of the inhibitoris retarded by at least 50% as compared to growth of non-contactedcells. More preferably, cell proliferation is inhibited by 100% (i.e.,the contacted cells do not increase in number). Most preferably, thephrase “inhibiting cell proliferation” includes a reduction in thenumber or size of contacted cells, as compared to non-contacted cells.Thus, a cleavage (e.g., hydrolyzation) product of a prodrug of aninhibitor of histone deacetylase according to the invention thatinhibits cell proliferation in a contacted cell may induce the contactedcell to undergo growth retardation, to undergo growth arrest, to undergoprogrammed cell death (i.e., to apoptose), or to undergo necrotic celldeath.

The cell proliferation inhibiting ability of the histone deacetylaseinhibitors according to the invention allows the synchronization of apopulation of asynchronously growing cells. For example, thehydrolzyation products of the prodrugs of histone deacetylase inhibitorsof the invention may be used to arrest a population of non-neoplasticcells grown in vitro in the G1 or G2 phase of the cell cycle. Suchsynchronization allows, for example, the identification of gene and/orgene products expressed during the G1 or G2 phase of the cell cycle.Such a synchronization of cultured cells may also be useful for testingthe efficacy of a new transfection protocol, where transfectionefficiency varies and is dependent upon the particular cell cycle phaseof the cell to be transfected. Use of the prodrugs of histonedeacetylase inhibitors of the invention allows the synchronization of apopulation of cells, thereby aiding detection of enhanced transfectionefficiency.

In some preferred embodiments, the contacted cell is a neoplastic cell.The term “neoplastic cell” is used to denote a cell that shows aberrantcell growth. Preferably, the aberrant cell growth of a neoplastic cellis increased cell growth. A neoplastic cell may be a hyperplastic cell,a cell that shows a lack of contact inhibition of growth in vitro, abenign tumor cell that is incapable of metastasis in vivo, or a cancercell that is capable of metastasis in vivo and that may recur afterattempted removal. The term “tumorigenesis” is used to denote theinduction of cell proliferation that leads to the development of aneoplastic growth. In some embodiments, the cleavage product of aprodrug of a histone deacetylase inhibitor of the invention induces celldifferentiation in the contacted cell. Thus, a neoplastic cell, whencontacted with a prodrug of an inhibitor of histone deacetylase of theinvention may be induced to differentiate, resulting in the productionof a daughter cell that is phylogenetic ally more advanced than thecontacted cell. In certain other preferred embodiments, the contactedcell is a fungal cell.

In some preferred embodiments, the contacted cell is in an animal Thus,the invention provides a method for treating a cell proliferativedisease or condition in an animal, or treating a fungal infection,comprising administering to an animal in need of such treatment atherapeutically effective amount of a prodrug of a histone deacetylaseinhibitor of the invention. Preferably, the animal is a mammal, morepreferably a domesticated mammal. Most preferably, the animal is ahuman.

The term “cell proliferative disease or condition” is meant to refer toany condition characterized by aberrant cell growth, preferablyabnormally increased cellular proliferation. Examples of such cellproliferative diseases or conditions include, but are not limited to,cancer, restenosis, and psoriasis. In particularly preferredembodiments, the invention provides a method for inhibiting neoplasticcell proliferation in an animal comprising administering to an animalhaving at least one neoplastic cell present in its body atherapeutically effective amount of a prodrug of a histone deacetylaseinhibitor of the invention.

It is contemplated that some cleavage products of the prodrugs of theinvention have inhibitory activity against a histone deacetylase from aprotozoal source. Thus, the invention also provides a method fortreating or preventing a protozoal disease or infection, comprisingadministering to an animal in need of such treatment a therapeuticallyeffective amount of a prodrug of a histone deacetylase inhibitor of theinvention. Preferably the animal is a mammal, more preferably a human.Preferably, the histone deacetylase inhibitor used according to thisembodiment of the invention inhibits a protozoal histone deacetylase toa greater extent than it inhibits mammalian histone deacetylases,particularly human histone deacetylases.

The present invention further provides a method for treating a fungaldisease or infection comprising administering to an animal in need ofsuch treatment a therapeutically effective amount of a prodrug of ahistone deacetylase inhibitor of the invention. Preferably the animal isa mammal, more preferably a human. Preferably, the histone deacetylaseinhibitor used according to this embodiment of the invention inhibits afungal histone deacetylase to a greater extent than it inhibitsmammalian histone deacetylases, particularly human histone deacetylases.

The term “therapeutically effective amount” is meant to denote a dosagesufficient to cause inhibition of histone deacetylase activity in thecells of the subject, or a dosage sufficient to inhibit cellproliferation or to induce cell differentiation in the subject.Administration may be by any route, including, without limitation,parenteral, oral, sublingual, transdermal, topical, intranasal,intratracheal, or intrarectal. In certain particularly preferredembodiments, prodrugs of the invention are administered intravenously ina hospital setting. In certain other preferred embodiments,administration may preferably be by the oral route.

When administered systemically, the prodrug of an histone deacetylaseinhibitor is preferably administered at a sufficient dosage to attain ablood level of the inhibitor from about 0.01 μM to about 100 μM, morepreferably from about 0.05 μM to about 50 μM, still more preferably fromabout 0.1 μM to about 25 μM, and still yet more preferably from about0.5 μM to about 25 μM. For localized administration, much lowerconcentrations than this may be effective, and much higherconcentrations may be tolerated. One of skill in the art will appreciatethat the dosage of histone deacetylase inhibitor necessary to produce atherapeutic effect may vary considerably depending on the tissue, organ,or the particular animal or patient to be treated.

In certain preferred embodiments of the third aspect of the invention,the method further comprises contacting the cell with an antisenseoligonucleotide that inhibits the expression of a histone deacetylase.The combined use of a nucleic acid level inhibitor (i.e., antisenseoligonucleotide) and a protein level inhibitor (i.e., inhibitor ofhistone deacetylase enzyme activity) results in an improved inhibitoryeffect, thereby reducing the amounts of the inhibitors required toobtain a given inhibitory effect as compared to the amounts necessarywhen either is used individually. Antisense oligonucleotides accordingto this aspect of the invention, when directed to mammalian HDAC, arecomplementary to regions of RNA or double-stranded DNA that encodeHDAC-1, HDAC-2, HDAC-3, HDAC-4, HDAC-5, HDAC-6, HDAC7, HDAC-8, HDAC-9,HDAC-10 and/or HDAC-11.

For purposes of the invention, the term “oligonucleotide” includespolymers of two or more deoxyribonucleosides, ribonucleosides, or2′-O-substituted ribonucleoside residues, or any combination thereof.Preferably, such oligonucleotides have from about 6 to about 100nucleoside residues, more preferably from about 8 to about 50 nucleosideresidues, and most preferably from about 12 to about 30 nucleosideresidues. The nucleoside residues may be coupled to each other by any ofthe numerous known internucleoside linkages. Such internucleosidelinkages include without limitation phosphorothioate,phosphorodithioate, alkylphosphonate, alkylphosphonothioate,phosphotriester, phosphoramidate, siloxane, carbonate,carboxymethylester, acetamidate, carbamate, thioether, bridgedphosphoramidate, bridged methylene phosphonate, bridged phosphorothioateand sulfone internucleoside linkages. In certain preferred embodiments,these internucleoside linkages may be phosphodiester, phosphotriester,phosphorothioate, or phosphoramidate linkages, or combinations thereof.The term oligonucleotide also encompasses such polymers havingchemically modified bases or sugars and/or having additionalsubstituents, including without limitation lipophilic groups,intercalating agents, diamines and adamantane. For purposes of theinvention the term “2′-O-substituted” means substitution of the 2′position of the pentose moiety with an —O-lower alkyl group containing1-6 saturated or unsaturated carbon atoms, or with an —O-aryl or allylgroup having 2-6 carbon atoms, wherein such alkyl, aryl or allyl groupmay be unsubstituted or may be substituted, e.g., with halo, hydroxy,trifluoromethyl, cyano, nitro, acyl, acyloxy, alkoxy, carboxyl,carbalkoxyl, or amino groups; or such 2′ substitution may be with ahydroxy group (to produce a ribonucleoside), an amino or a halo group,but not with a 2′-H group. The term “oligonucleotide” also encompasseslinked nucleic acid and peptide nucleic acid.

Particularly preferred antisense oligonucleotides utilized in thisaspect of the invention include chimeric oligonucleotides and hybridoligonucleotides.

For purposes of the invention, a “chimeric oligonucleotide” refers to anoligonucleotide having more than one type of internucleoside linkage.One preferred example of such a chimeric oligonucleotide is a chimericoligonucleotide comprising a phosphorothioate, phosphodiester orphosphorodithioate region, preferably comprising from about 2 to about12 nucleotides, and an alkylphosphonate or alkylphosphonothioate region(see e.g., Pederson et al. U.S. Pat. Nos. 5,635,377 and 5,366,878).Preferably, such chimeric oligonucleotides contain at least threeconsecutive internucleoside linkages selected from phosphodiester andphosphorothioate linkages, or combinations thereof.

For purposes of the invention, a “hybrid oligonucleotide” refers to anoligonucleotide having more than one type of nucleoside. One preferredexample of such a hybrid oligonucleotide comprises a ribonucleotide or2′-O-substituted ribonucleotide region, preferably comprising from about2 to about 12 2′-O-substituted nucleotides, and a deoxyribonucleotideregion. Preferably, such a hybrid oligonucleotide will contain at leastthree consecutive deoxyribonucleosides and will also containribonucleosides, 2′-O-substituted ribonucleosides, or combinationsthereof (see e.g., Metelev and Agrawal, U.S. Pat. No. 5,652,355).

The exact nucleotide sequence and chemical structure of an antisenseoligonucleotide utilized in the invention can be varied, so long as theoligonucleotide retains its ability to inhibit expression of the gene ofinterest. This is readily determined by testing whether the particularantisense oligonucleotide is active by quantitating the mRNA encoding aproduct of the gene, or in a Western blotting analysis assay for theproduct of the gene, or in an activity assay for an enzymatically activegene product, or in a soft agar growth assay, or in a reporter geneconstruct assay, or an in vivo tumor growth assay, all of which aredescribed in detail in this specification or in Ramchandani et al.(1997) Proc. Natl. Acad. Sci. USA 94: 684-689.

Antisense oligonucleotides utilized in the invention may conveniently besynthesized on a suitable solid support using well known chemicalapproaches, including H-phosphonate chemistry, phosphoramiditechemistry, or a combination of H-phosphonate chemistry andphosphoramidite chemistry (i.e., H-phosphonate chemistry for some cyclesand phosphoramidite chemistry for other cycles). Suitable solid supportsinclude any of the standard solid supports used for solid phaseoligonucleotide synthesis, such as controlled-pore glass (CPG) (see,e.g., Pon, R.T. (1993) Methods in Molec. Biol. 20: 465-496).

Particularly, preferred oligonucleotides have nucleotide sequences offrom about 13 to about 35 nucleotides which include the nucleotidesequences shown in Tables 1-3. Yet additional particularly preferredoligonucleotides have nucleotide sequences of from about 15 to about 26nucleotides of the nucleotide sequences shown in Tables 1-3.

TABLE 1 SEQ ID TARGET NO. SEQUENCE (**) 15′-GAG ACA GCA GCA CCA GCG GG-3′ 17-36 25′-ATG ACC GAG TGG GAG ACA GC-3′ 21-49 35′-GGA TGA CCG AGT GGG AGA CA-3′ 31-50 45′-CAG GAT GAC CGA GTG GGA GA-3′ 33-52 55′-TGT GTT CTC AGG ATG ACC GA-3′ 41-60 65′-GAG TGA CAG AGA CGC TCA GG-3′ 62-81 75′-TTC TGG CTT CTC CTC CTT GG-3′ 1504-1523 85′-CTT GAC CTC CTC CTT GAC CC-3′ 1531-1550 95′-GGA AGC CAG AGC TGG AGA GG-3′ 1565-1584 105′-GAA ACG TGA GGG ACT CAG CA-3′ 1585-1604 115′-CCG TCG TAG TAG TAA CAG ACT 138-160 TT-3′ 125′-TGT CCA TAA TAG TAA TTT CCA A-3′ 166-187 135′-CAG CAA ATT ATG AGT CAT GCG GAT 211-236 TC-3′ (**) target referencenumbering is in accordance with HDAC-1, GenBank Accession Number U50079.

TABLE 2 SEQ ID TARGET NO. SEQUENCE (***) 145′-CTC CTT GAC TGT ACG CCA TG-3′  1-20 155′-TGC TGC TGC TGC TGC TGC CG-3′ 121-141 165′-CCT CCT GCT GCT GCT GCT GC-3′ 132-152 175′-CCG TCG TAG TAG TAG CAG ACT TT- 138-160 3′ 185′-TGT CCA TAA TAA TAA TTT CCA A-3′ 166-187 195′-CAG CAA GTT ATG GGT CAT GCG GAT 211-236 TC-3′ 205′-GGT TCC TTT GGT ATC TGT TT-3′ 1605-1625 (***) target referencenumbering is in accordance with HDAC-2, GenBank Accession Number U31814.

TABLE 3 SEQ ID TARGET NO. SEQUENCE (***) 215′-GCT GCC TGC CGT GCC CAC CC-3′ 514-533 (***) target referencenumbering is in accordance with HDAC-4

The following examples are intended to further illustrate certainpreferred embodiments of the invention, and are not intended to limitthe scope of the invention.

EXAMPLES Preparation of Amines Methyl-3-aminophenylacetate (1)

To a solution of 3-aminophenylacetic acid (3 g, 19.85 mmol) in methanol(50 mL) at room temperature was added HCl conc. (37%, 7.5 mL). Themixture was stirred 6 h at room temperature then treated with asaturated aqueous solution of NaHCO₃. The solvent was removed underreduced pressure then the aqueous phase was extracted several times withCH₂Cl₂. The combined organic extracts were dried over (MgSO₄) andevaporated. The crude mixture was purified by flash chromatography usinghexane/AcOEt (1:1) yielding 1 as a yellow oil (3.06 g, 79%).

¹H NMR: (300 MHz, CDCl₃): δ 7.10 (t, J=8 Hz, 1H), 6.68-6.58 (m, 3H),3.69-3.65 (m, 5H), 3.53 (s, 2H).

Methyl-4-aminophenyl benzoate (2)

To a solution of 4-aminobenzoic acid (10 g, 72.92 mmol) in methanol (200mL) at room temperature was added HCl conc. (37%, 25 mL). The solutionmixture was heated overnight at 70° C. Once the solution was clear(completed) the reaction was treated with a saturated aqueous solutionof NaHCO₃ and Na₂CO₃ powder until pH 9. The solvent was then evaporatedunder reduced pressure and the aqueous phase was extracted several timeswith AcOEt. The combined organic extracts were dried over (MgSO₄) andevaporated. The crude product 2 (9.30 g 85%) was obtained as a beigesolid and was clean enough to use without further purification.

¹H NMR: (300 MHz, CDCl₃): δ 7.85 (d, J=8 Hz, 2H), 6.63 (d, J=8 Hz, 2H),4.04 (broad s. 2H), 3.85 (s. 3H).

Methyl-4-aminophenylacetate (3)

To a solution of 4-aminophenylacetic acid (10 g, 66 2 mmol) in methanol(150 mL) at room temperature was added HCl conc. (37% 25 mL). Themixture became yellow and was stirred overnight. The reaction mixturewas then quenched with a saturated aqueous solution of NaHCO₃. Themethanol was evaporated under reduced pressure and the aqueous layer wasextracted several times with AcOEt. The combined organic extracts weredried over (MgSO₄) and evaporated. The crude residue was purified byflash chromatography using hexane/AcOEt (4:1) as solvent mixtureyielding 3 as a yellow oil (9.44 g, 74%).

¹H NMR: (300 MHz, CDCl₃): δ 7.05 (d, J=10 Hz, 2H), 6.65 (d, J=10 Hz,2H), 3.65 (s, 3H), 3.63 (broad s, 2H), 3.51 (s, 2H).

Example 12-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-N-hydroxy-acetamide (4)

Step 1: Methyl-2-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-acetate(5)

To a solution of 3 (500 mg, 2.56 mmol), in CH₂Cl₂ (8 mL) at roomtemperature were added Et₃N (712 μμL, 5.12 mmol) followed by2-benzothiophenesulfonyl chloride (712 mg, 3.07 mmol). The mixture wasstirred overnight at room temperature then quenched with a saturatedaqueous solution of NaHCO₃. The phases were separated and the aqueouslayer was extracted several times with CH₂Cl₂. The combined organicextracts were dried over (MgSO₄) and evaporated. The mixture of the monoand bis alkylated products were dissolved in methanol (˜8 mL) and NaOMewas added (691 mg, 12.8 mmol). The resulting mixture was heated at 60°C. for 30 min the HCl 1N was added until pH 2. Then a saturated aqueoussolution of NaHCO₃ was added until pH 7-8. The solvent was evaporatedunder reduced pressure then the aqueous layer was extracted severaltimes with CH₂Cl₂. The combined organic extracts were dried over (MgSO₄)and evaporated. The residue was purified by flash chromatography usingtoluene/AcOEt 7:3 as solvent mixture and a second flash chromatographyusing CH₂Cl₂/acetone 98:2 as solvent yielding the title compound 5 asyellowish powder (487 mg, 53%).

¹H NMR: (300 MHz, CDCl₃): δ 7.80 (d, J=8 Hz, 2H), 7.75 (s, 1H), 7.44 (m,2H), 7.14 (m, 4H), 6.79 (broad s, 1H) 3.67 (s, 3H), 3.56 (s, 2H)

Step 2: 2-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-acetic acid (6)

To a solution of 5 from step 1 (451 mg, 1.25 mmol) in a solvent mixtureof THF (20 mL) and H₂O (20 mL) at room temperature was added LiOH (524mg, 12.5 mmol). The mixture was stirred for 2 h at room temperature andthen was treated with a saturated aqueous solution of NH₄Cl. Theresulting solution was extracted several times with AcOEt. The combinedorganic extracts were dried over (MgSO₄). The crude residue was thenpurified by flash chromatography using CH₂Cl₂/MeOH (9:1) as solventmixture yielding the title compound 6 as white solid (404 mg, 93%).

¹H NMR: (300 MHz, DMSO-d₆): δ 8.03 (d, J=8 Hz, 1H), 7.97 (d, J=7 Hz,1H), 7.92 (s, 1H), 7.50-7.45 (m, 2H), 7.13-7.06 (m, 4H), 3.44 (s, 2H).

Step 3:2-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-N-hydroxy-acetamide (4)

Method A:

To a solution of 6 (150 mg, 0.432 mmol) in a solvent mixture of CH₂Cl₂(10 mL) and THF (5 mL) was added at room temperature1,3-dicyclohexylcarbodiimide (DCC, 116 mg, 0.563 mmol). The reactionmixture was stirred 30 min at room temperature then NH₂OTHP (76 mg,0.650 mmol) and dimethylaminopyridine (DMAP, 5 mg) were added. Thesolution was stirred over night at room temperature and the solventswere evaporated under reduced pressure. The crude material was purifiedby flash chromatography using CH₂Cl₂/MeOH (9:1) as solvent. The residuewas dissolved in MeOH (−10 mL) and 10-camphorsulfonic acid (CSA, 100 mg,0.432 mmol) was added. The mixture was stirred at room temperatureovernight then treated with a saturated aqueous solution of NaHCO₃. Thesolvent was evaporated under reduced pressure and the aqueous phase wasextracted several times with CH₂Cl₂ (3×) and AcOEt (3×). The combinedorganic extracts were dried over (MgSO₄) and evaporated. The crudeproduct was purified by preparative high pressure liquid chromatographyon reversed phase silica gel using a gradient of water/CH₃CN (10-65%)yielding the title compound 4 as yellowish solid (70 mg, 45%).

¹H NMR (300 MHz, CD₃OD): δ 7.92-7.88 (m, 2H), 7.80 (s, 1H), 7.50-7.45(m, 2H), 7.23-7.16 (m, 4H) 3.35 (s, 2H).

Except where otherwise indicated, the following compounds were preparedby procedures analogous to those described in Example 1, butsubstituting the sulfonyl chloride indicated for2-benzothiophenesulfonyl chloride in step 1.

Example 2 2-[4-(2-nitrobenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide(7)

Sulfonyl chloride: 2-nitrobenzenesulfonyl chloride

Yield: Step 1: 82%

Yield: Step 2: 99%

Yield: Step 3: 19%

¹H NMR (300 MHz, DMSO-d₆); δ 10.59 (s, 1H); 8.78 (s, 1H); 7.94 (s, 2H),7.81 (s, 2H), 7.20-7.02 (m, 4H); 3.13 (s, 2H).

Example 32-[4-(2.5-dichlorobenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide (8)

Sulfonyl chloride: 2.5-Dichlorobenzenesulfonyl chloride

Yield: Step 1: 66%

Yield: Step 2: 96%

Yield: Step 3: 66%

¹H NMR (300 MHz, DMSO-d₆); δ 10.68 (s, 1H), 8.88 (s, 1H), 7.95 (s, 1H),7.67 (s, 2H); 7.13 (d, 2H, J=8 Hz 7.02 (d, 2H, J=8 Hz 3.16 (s, 2H)

Example 42-[4-(4-methylbenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide (9)

Sulfonyl chloride: 4-methylbenzenesulfonyl chloride Step 1: Yield 100%Step 2: 2-[4-(4-methylbenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide(9) Method B:

To a solution ofmethyl-2-[4-(4-methylbenzenesulfonylamino)]phenylacetate (459 mg, 1.44mmol) in methanol (10 mL), at room temperature were added hydroxylaminehydrochloride (200 mg, 2.88 mmol) followed by sodium methoxide (389 mg,7.19 mmol). The resulting mixture was heated overnight at 60° C. thentreated with HCl (1N) until pH 2. The solvent was evaporated underreduced pressure then the aqueous phase was extracted several times withCH₂Cl₂. The combined organic extracts were dried over (MgSO₄) thenevaporated. The crude mixture was purified by flash chromatography usingCH₂Cl₂/MeOH (9:1) as solvent mixture yielding the title compound 9 (244mg, 53%) as a white powder.

¹H NMR (300 MHz, acetone-d₆); δ 7.68 (d, J=8 Hz, 2H); 7.29 (d, J=8 Hz,2H), 7.15 (br. s, 4H), 3.33 (s, 2H, CH₂), 2.33 (s, 3H, CH₃).

The following compounds were prepared following procedures analogous tothose described in Example 1, step 1, and Example 4, step 2 (Method B),but substituting the sulfonyl chloride indicated for2-benzothiophenesulfonyl chloride in step 1.

Example 5 2-[4-(3-trifluoromethylbenzenesulfonylamino)-phenyl]-N-hydroxyacetamide (10)

Sulfonyl chloride: 3-trifluoromethylbenzenesulfonyl chloride

Yield: Step 1: 70%

Yield: Step 2: 49%

¹H NMR (300 MHz, acetone-d₆); δ=8.09 (s, 1H), 8.05 (d, 1H, J=8 Hz), 7.95(d, 1H, J=8 Hz); 7.77 (t, 1H, J=8 Hz); 7.21 (d, 2H, J=8 Hz), 7.13 (d,2H, J=8 Hz); 3.35 (s, 2H, CH₂)

Example 6 2-[4-(tert-butylsulfonylamino)-phenyl]-N-hydroxy-acetamide(11)

Sulfonyl chloride: 4-tert-butylsulfonyl chloride

Yield: Step 1: 76%

Yield: Step 2: 40%

¹H NMR (300 MHz, acetone-d₆); δ 7.75 (d, 2H, J=9 Hz), 7.56 (d, 2H, J=9Hz); 7.17 (s, 4H); 3.34 (s, 2H), 1.29 (s, 9H)

The following compound was prepared following procedures analogous tothose described in Example 1, steps 1-2, substituting the sulfonylchloride indicated for 2-benzothiophenesulfonyl chloride in step 1,followed by hydroxamic acid formation using

Method C Example 72-[2-(naphthylsulfonylamino)-phenyl]-N-hydroxy-acetamide (12)

Sulfonyl chloride: 2-naphthylsulfonyl chloride

Yield: Step 1: 100%

Yield: Step 2: 100%

Step 3: 2-[2-(naphthylsulfonylamino)-phenyl]-N-hydroxy-acetamide (12)Method C:

To a solution of 2-[2-(naphthylsulfonylamino)]-phenylacetic acid (191mg, 0.563 mmol) in CH₂Cl₂ (20 mL) at room temperature were added DMF (5drop) followed by (COCl)₂ (250 μL, 2.81 mmol). The mixture became yellowand solidification appeared. The reaction was stirred 90 min at roomtemperature then (COCl)₂ was added until no bubbling (˜1 mL). Then thesolvents were evaporated under reduced pressure. The crude material wasdissolved in CH₂Cl₂ and TMSONH₂ (3 mL) was added to the solution. Thereaction was exothermic and the resulting mixture was stirred 2 h atroom temperature then treated with HCl (1N) until pH 2. The phases wereseparated and the aqueous layer was extracted several times with CH₂Cl₂.The combined organic extracts were dried over (MgSO₄) then evaporated.The crude compound was purified 3 times by flash chromatography usingCH₂Cl₂/MeOH (9:1) as solvent mixture then another purification usingpreparative high pressure liquid chromatography using reversed phasechromatography with a gradient of water/CH₃CN (10-70%) yielding thetitle compound 12 as a white powder (29 mg, 15%).

¹H NMR (300 MHz, acetone-d₆); δ 9.13 (s, 1H), 8.42 (s, 1H), 8.08-7.97(m, 3H), 7.82 (dd, 1H, J=9 Hz, 1.5 Hz), 7.70-7.63 (m, 2H), 7.21-7.14 (m,4H), 3.50 (s, 2H)

The following compound was prepared following procedures analogous tothose described in Example 1, steps 1-2, substituting the indicatedsulfonyl chloride and amine indicated for 2-benzothiophenesulfonylchloride and 3 in step 1, followed by hydroxamic acid formation usingMethod D.

Example 8N-hydroxy-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-benzamide (13)

Sulfonyl chloride: 2-Benzothiophenesulfonyl chloride

Amine: Methyl-4-aminobenzoate (2)

Yield: Step 1: 80%

Yield: Step 2: 69%

Step 3:N-hydroxy-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-benzamide (13)Method D:

To a solution of 2-[4-benzo[b]thiophene-2-sulfonylamino]benzoic acid(300 mg, 0.90 mmol) in DMF (20 mL) at room temperature were added1-(3-dimethyl-aminopropyl)-3-ethylcarbodiimide hydrochloride (EDC, 207mg, 1.08 mmol), and 1-Hydroxybenzotriazole hydrate (HOBT, 182 mg, 1.35mmol). The mixture was stirred 20 min. at room temperature then NH₂OTHP(158 mg, 1.35 mmol) was added. The resulting mixture was heated at 50°C. for 24 h then stirred at room temperature for 24 h. The DMF solventwas evaporated under reduced pressure and the residue was dissolved inCH₂Cl₂ and washed with brine or a saturated aqueous solution of NaHCO₃.The combined organic extracts were dried over (MgSO₄) then condensed.The crude compound was purified by flash chromatography usingCH₂Cl₂/MeOH (9:1) as solvent mixture. The residue was then dissolved inmethanol (20 mL) then 10-camphorsulfonic acid (CSA, 100 mg, 0.45 mmol)was added. The mixture was stirred 2 h at room temperature then thesolvents were evaporated under reduced pressure at room temperature toavoid thermal decomposition. The crude was purified by flashchromatography using CH₂Cl₂/MeOH (9:1) as solvent mixture. A secondpurification was performed using a preparative high pressure liquidchromatography using a gradient of water/CH₃CN (10-85%) as solventgiving the title compound 13 as a red solid (212 mg, 68%).

¹H NMR (300 MHz, acetone-d₆); δ 10.69 (s, 1H), 9.70 (s, 1H); 8.01-7.97(m, 3H), 7.77 (d, 2H, J=9 Hz); 7.55-7.39 (m, 4H).

Example 92-[3-benzo[b]thiopene-2-sulfonylamino)-phenyl]N-hydroxy-acetamide (14)

Sulfonyl chloride: 2-Benzothiophenesulfonyl chloride

Amine: Methyl-3-aminophenyl acetate (1)

Yield: Step 1: 88%

Yield: Step 2: 89%

Yield: Step 3: 32%

¹H NMR (300 MHz, Acetoned₆); δ 10.20 (s, 1H), 8.33 (s, 1H), 7.99-7.95(m, 3H), 7.53-7.43 (m, 2H), 7.35 (s, 1H), 7.21-7.17 (m, 2H), 7.06-7.03(m, 1H), 3.38 (s, 2H)

Example 102-[4-(3,4-dichlorobenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide (15)

Sulfonyl chloride: 3.4-Dichlorobenzenesulfonyl chloride

Yield: Step 1: 80%

Yield: Step 2: 67%

Yield: Step 3: 81%

¹H NMR (300 MHz, acetone-d₆); δ 10.12 (s, 1H), 9.15 (s, 1H), 7.92 (s,1H), 7.74-7.71 (m, 2H), 7.23 (d, 2H, J=9 Hz), 7.14 (d, 2H, J=9 Hz), 3.36(s, 2H)

Example 11 2-[4-(2-Thiophenesulfonylamino)-phenyl]-N-hydroxy-acetamide(16)

Sulfonyl chloride: 2-Thiophenesulfonyl chloride

Yield: Step 1: 84%

Yield: Step 2: 83%

Yield: Step 3: 9%

¹H NMR (300 MHz, acetone-d₆); δ 7.78 (s, 1H), 7.53 (s, 1H), 7.21 (s,4H), 7.09 (s, 1H), 3.37 (s, 2H)

Example 122-[4-(3-nitrobenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide (17)

Sulfonyl chloride: 3-Nitrobenzenesulfonyl chloride

Yield: Step 1: 47%

Yield: Step 2: 34%

Yield: Step 3: 16%

¹H NMR (300 MHz, acetone-d₆); δ 9.31 (s, 1H), 8.59 (s, 1H), 8.45 (d, 1H,J=8 Hz), 8.16 (d, 1H, J=8 Hz), 7.85 (t, 1H, J=8 Hz), 7.20-7.14 (m, 4H),3.35 (s, 2H)

Example 13 2-[4-(8-quinolinesulfonylamino)-phenyl]-N-hydroxy-acetamide(18)

Sulfonyl chloride: 8-quinolinesulfonyl chloride

Yield: Step 1: 83%

Yield: Step 2: 78%

Yield: Step 3: 42%

¹H NMR (300 MHz, acetone-d₆); δ 9.17 (s, 1H), 8.50 (d, 1H, J=8 Hz), 8.33(d, 1H, J=8 Hz), 8.21 (d, 1H, J=8 Hz), 7.71-7.68 (m, 3H), 7.05 (broads., 4H), 3.22 (s, 2H)

Example 142-[4-(4-bromobenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide (19)

Sulfonyl chloride: 4-Bromobenzenesulfonyl chloride

Yield: Step 1: 80%

Yield: Step 2: 81%

Yield: Step 3: 48%

¹H NMR (300 MHz, acetone-d₆); δ 9.17 (s, 1H), 7.72 (s, 4H), 7.19-7.14(m, 4H), 3.35 (s, 2H)

Example 15 N-Hydroxy-5-[3-benzenesulfonylamino)-phenyl]-pentanamide (26)

Step 1: 3-(benzenesulfonylamino)-phenyl iodide (21)

To a solution of 3-iodoaniline (5 g, 22 8 mmol), in CH₂Cl₂ (100 mL),were added at room temperature Et₃N (6.97 mL) followed bybenzenesulfonyl chloride (5.84 mL). The mixture was stirred 4 h then awhite precipitate was formed. A saturated aqueous solution of NaHCO₃ wasadded and the phases were separated. The aqueous layer was extractedseveral times with CH₂Cl₂ and the combined extracts were dried over(MgSO₄) then evaporated. The crude mixture was dissolved in MeOH (100mL) and NaOMe (6 g), was added and the mixture was heated 1 h at 60° C.The solution became clear with time and HCl (1N) was added. The solventwas evaporated under reduced pressure then the aqueous phase wasextracted several times with CH₂Cl₂. The combined organic extracts weredried over (MgSO₄) and evaporated. The crude material was purified byflash chromatography using (100% CH₂Cl₂) as solvent yielding the titlecompound 21 (7.68 g, 94%) as yellow solid.

¹H NMR: (300 MHz, CDCl₃): δ 7.82-7.78 (m, 2H), 7.60-7.55 (m, 1H),7.50-7.42 (m, 4H), 7.10-7.06 (m, 1H), 6.96 (t, J=8 Hz, 1H), 6.87 (broads, 1H).

Step 2: 3-(benzenesulfonylamino)-phenyl-propargylic alcohol (22)

To a solution of 21 (500 mg, 1.39 mmol) in pyrrolidine (5 mL) at roomtemperature was added Pd(PPh₃)₄ (80 mg, 0.069 mmol), followed by CuI (26mg, 0.139 mmol). The mixture was stirred until complete dissolution.Propargylic alcohol (162 μL, 2.78 mmol) was added and stirred 6 h atroom temperature. Then the solution was treated with a saturated aqueoussolution of NH₄Cl and extracted several times with AcOEt. The combinedorganic extracts were dried over (MgSO₄) then evaporated. The residuewas purified by flash chromatography using hexane/AcOEt (1:1) as solventmixture yielding 22 (395 mg, 99%) as yellow solid.

¹H NMR: (300 MHz, CDCl₃): δ 7.79-7.76 (m, 2H), 7.55-7.52 (m, 1H), 7.45(t, J=8 Hz, 2H), 7.19-7.15 (m, 3H), 7.07-7.03 (m, 1H), 4.47 (s, 2H).

Step 3: 5-[3-(benzenesulfonylamino)-phenyl]-4-yn-2-pentenoate (23)

To a solution of 22 (2.75 g, 9.58 mmol) in CH₃CN (150 mL) at roomtemperature were added 4-methylmorpholine N-oxide (NMO, 1.68 g, 14.37mmol) followed by tetrapropylammonium perruthenate (TPAP, 336 mg, 0.958mmol). The mixture was stirred at room temperature 3 h, and thenfiltrated through a Celite pad with a fritted glass funnel. To thefiltrate carbethoxymethylenetriphenyl-phosphorane (6.66 g, 19.16 mmol)was added and the resulting solution was stirred 3 h at roomtemperature. The solvent was evaporated and the residue was dissolved inCH₂Cl₂ and washed with a saturated aqueous solution of NH₄Cl. Theaqueous layer was extracted several times with CH₂Cl₂ then the combinedorganic extract were dried over (MgSO₄) and evaporated. The crudematerial was purified by flash chromatography using hexane/AcOEt (1:1)as solvent mixture giving 23 (1.21 g, 36%) as yellow oil.

¹H NMR: (300 MHz, CDCl₃): δ 7.81 (d, J=8 Hz, 2H), 7.56-7.43 (m, 3H),7.26-7.21 (m, 3H), 7.13-7.11 (m, 1H), 6.93 (d, J=16 Hz, 1H), 6.29 (d,J=16 Hz, 1H), 4.24 (q, J=7 Hz, 2H), 1.31 (t, J=7 Hz, 3H).

Step 4: 5-[3-(benzenesulfonylamino)-phenyl]-4-yn-2-pentenic acid (24)

To a solution of 23 (888 mg, 2.50 mmol) in a solvent mixture of THF (10mL) and water (10 mL) at room temperature was added LiOH (1.04 g, 25.01mmol). The resulting mixture was heated 2 h at 60° C. and treated withHCl (1N) until pH 2. The phases were separated and the aqueous layer wasextracted several times with AcOEt. The combined organic extracts weredried over (MgSO₄) then evaporated. The crude residue was purified byflash chromatography using CH₂Cl₂/MeOH (9:1) as solvent mixture yielding24 (712 mg, 88%), as white solid.

¹H NMR: (300 MHz, DMSO-d₆): δ 7.78-7.76 (m, 2H), 7.75-7.53 (m, 3H),7.33-7.27 (m, 1H), 7.19-7.16 (m, 3H), 6.89 (d, J=16 Hz, 1H), 6.33 (d,J=16 Hz, 1H).

Step 5: 5-[3-(benzenesulfonylamino)-phenyl]-pentanoic acid (25)

To a solution 24 (100 mg, 0.306 mmol), in MeOH (6 mL) at roomtemperature was added a solution of Pd/C (10%, 20 mg, 1 mL MeOH). Thereaction mixture was degassed and purged several times with H₂ gas witha final pressure of 60 psi. The mixture was stirred 2 h at roomtemperature then the resulting solution was filtrated over a silica gelpad with a fritted glass funnel. The solvent was evaporated yielding 25(68 mg, 96%) and it was used directly for the next step without furtherpurification.

¹H NMR: (300 MHz, acetone-d₆): δ 7.81-7.78 (m, 2H), 7.56-7.46 (m, 3H),7.11-7.01 (m, 3H), 6.87 (d, J=8 Hz, 1H), 2.49 (broad s, 2H), 2.25 (broads, 2H), 1.52 (broad s, 4H)

Step 6: N-Hydroxy-5-[3-benzenesulfonylamino)-phenyl]-pentanamide (26)

To a solution of 25 (100 mg, 300 mmol) in DMF (10 mL) at roomtemperature were added 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimidehydrochloride (EDC, 69 mg, 0.320 mmol), and 1-hydroxybenzotriazolehydrate (HOBT, 61 mg, 0.45 mmol). The mixture was stirred 20 min. atroom temperature then NH₂OTHP (53 mg, 0.45 mmol) was added. Theresulting mixture was heated overnight at 50° C. The DMF solvent wasevaporated under reduced pressure and the residue was dissolved inCH₂Cl₂ and washed with brine or a saturated aqueous solution of NaHCO₃.The combined organic extracts were dried over (MgSO₄) then evaporated.The crude compound was purified by flash chromatography usinghexane/acetone (7:3) as solvent mixture. The residue was then dissolvedin MeOH (20 mL) then 10-camphorsulfonic acid (CSA, 35 mg, 150 mmol) wasadded. The mixture was stirred 2 h at room temperature then the solventswere evaporated under reduced pressure at room temperature to avoidthermal decomposition. The crude mixture was purified by flashchromatography using CH₂Cl₂/MeOH (9:1) as solvent mixture giving 26 as ayellowish solid (62 mg, 60%).

¹H NMR: (300 MHz, acetone-d₆): δ=7.80-7.78 (m, 2H), 7.56-7.52 (m, 3H),7.13-6.89 (m, 4H), 2.52 (broad s, 2H), 2.10 (broad s, 2H), 1.53 (broads, 4H)

Example 16N-Hydroxy-5-[4-(benzenesulfonylamino)-phenyl]-4-yn-2-pentanamide (32)

Step 1: 4-(benzenesulfonylamino)-phenyl iodide (28)

Compound 28 was prepared using the procedure described in Example 15,step 1, but substituting 4-iodoaniline for 3-iodoaniline.

Yield: 97%

¹H NMR: (300 MHz, CDCl₃): δ 9.15 (broad s, 1H), 7.82 (d, J=8 Hz, 2H),7.68-7.51 (m, 5H), 7.05 (d, J=8 Hz, 2H).

Step 2: 4-(benzenesulfonylamino)-phenyl-propargylic alcohol (29)

Compound 29 was prepared using the procedure described in Example 15,step 2 but substituting compound 21 for compound 28.

Yield: 61%

¹H NMR: (300 MHz, acetone-d₆): δ 7.83-7.80 (m, 2H), 7.62-7.51 (m, 3H),7.30 (d, J=8 Hz, 2H), 7.21 (d, J=8 Hz, 2H), 4.36 (s, 2H), 2.80 (broad s,2H).

Step 3: 5-[4-(benzenesulfonylamino)-phenyl]-4-yn-2-pentenoate (30)

Compound 30 was prepared using the procedure described in Example 15,step 3 but substituting compound 22 for compound 29

Yield: 16%

¹H NMR: (300 MHz, CDCl₃): δ 7.81-7.78 (m, 2H), 7.59-7.43 (m, 3H), 7.34(d, J=8 Hz, 2H), 7.05 (d, J=8 Hz, 2H), 6.93 (d, J=16 Hz, 1H), 6.26 (d,J=16 Hz, 1H), 4.23 (q, J=7 Hz, 2H), 1.30 (t, J=7 Hz, 3H).

Step 4: 5-[4-(benzenesulfonylamino)-phenyl]-4-yn-2-pentenic acid (31)

Compound 31 was prepared using the procedure described in Example 15step 4 but substituting compound 23 for compound 30

Yield: 92%

¹H NMR: (300 MHz, acetone-d₆): δ 7.87-7.84 (m, 2H), 7.62 (m, 3H), 7.42(d, J=8 Hz, 2H), 7.28 (d, J=8 Hz, 2H), 6.94 (d, J=16 Hz, 1H), 6.29 (d,J=16 Hz, 1H).

Step 5: N-hydroxy-5-[4-(benzenesulfonylamino)-phenyl]-4-yn-2-pentanamide(32)

Compound 32 was prepared using the procedure described in Example 15step 6 but substituting compound 25 for compound 31

Yield: 78%

¹H NMR: (300 MHz, acetone-d₆): δ 7.84 (broad s, 2H), 7.60-7.55 (m, 3H),7.38-7.30 (m, 4H), 6.84 (d, J=16 Hz, 1H), 6.40 (d, J=16 Hz, 1H).

Example 17 N-Hydroxy-5-[4-benzenesulfonylamino)-phenyl]-pentanamide (34)Step 1: 5-[4-(benzenesulfonylamino)-phenyl]-pentanoic acid (33)

Compound 33 was prepared using the procedure described in Example 15step 5 but substituting compound 24 for compound 31.

Yield: 100%

¹H NMR: (300 MHz, acetone-d₆): δ=7.78-7.75 (m, 2H), 7.56-7.46 (m, 3H),7.16-7.05 (m, 4H), 2.52 (broad s, 2H), 2.29-2.25 (m, 2H), 1.56 (broad s,4H).

Step 2: N-Hydroxy-5-[4-benzenesulfonylamino)-phenyl]-pentanamide (34)

Compound 34 was prepared using the procedure described in Example 15step 6 but substituting compound 25 for compound 33.

Yield: 62%

¹H NMR: (300 MHz, acetone-d₆): δ 7.78-7.75 (m, 2H), 7.59-7.51 (m, 3H),7.09 (broad s, 4H), 2.85 (broad s, 1H), 2.53 (broad s, 2H), 2.05 (broads, 2H), 1.56 (broad s, 4H).

Example 18 N-Hydroxy-3-[4-(benzenesulfonylamino)-phenyl]-2-propenamide(36)

Step 1: 3-[4-(benzenesulfonylamino)-phenyl]-2-propenoic acid (35)

To a solution of 28 (500 mg, 1.39 mmol), in DMF (10 mL) at roomtemperature were added tris(dibenzylideneacetone)dipalladium(0)(Pd₂(dba)₃; 38 mg, 1.67 mmol), tri-o-tolylphosphine (P(o-tol)₃, 25 mg,0.83 mmol), Et₃N (483 μμL, 3.48 mmol) and finally acrylic acid (84 μμL,1.67 mmol). The resulting solution was degassed and purged several timeswith N₂ then heated overnight at 100° C. The solution was filtratedthrough a Celite pad with a fritted glass funnel then the filtrate wasevaporated. The residue was purified by flash chromatography usingCH₂Cl₂/MeOH (95:5) as solvent mixture yielding the title compound 35(415 mg, 99%) as yellowish solid.

¹H NMR: (300 MHz, acetone-d₆): δ 7.88-7.85 (m, 2H), 7.62-7.55 (m, 6H),7.29 (d, J=9 Hz, 2H), 6.41 (d, J=16 Hz, 1H), 2.95 (s, 1H), 2.79 (s, 1H).

Step 2: N-Hydroxy-3-[4-(benzenesulfonylamino)-phenyl]-2-propenamide (36)

To a solution of 35 (200 mg, 0.660 mmol) in DMF (10 mL) at roomtemperature were added 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimidehydrochloride (EDCI, 151 mg, 0.79 mmol), and 1-Hydroxybenzotriazolehydrate (HOBT, 134 mg, 0.99 mmol). The mixture was stirred 20 min. atroom temperature then NH₂OTHP (116 mg, 0.99 mmol) was added. Theresulting mixture was heated at 50° C. for 24 h then the DMF solvent wasevaporated under reduced pressure and the residue was dissolved inCH₂Cl₂, washed with a saturated aqueous solution of NaHCO₃. The combinedorganic extracts were dried over (MgSO₄) then condensed. The crudecompound was purified by flash chromatography using Hexane/acetone (7:3)as solvent mixture. The residue was then dissolved in MeOH (10 mL) then10-camphorsulfonic acid (CSA, 77 mg, 0.33 mmol) was added. The mixturewas stirred 2 h at room temperature then the solvents were evaporatedunder reduced pressure at room temperature to avoid thermaldecomposition. The crude product was purified by flash chromatographyusing CH₂Cl₂/MeOH (9:1) as solvent mixture giving compound 36 (116 mg,55%) as a orange solid.

¹H NMR: (300 MHz, acetone-d₆): δ 7.85-7.83 (m, 2H), 7.64-7.47 (m, 6H),7.26 (d, J=8 Hz, 2H), 6.48 (m, 1H), 2.82 (s, 1H), 2.79 (s, 1H).

Example 19 N-Hydroxy-3-[4-(benzenesulfonylamino)-phenyl]-2-propanamide(38) Step 1: 3-[4-(benzenesulfonylamino)-phenyl]-2-propionic acid (37)

To a solution of 35 (350 mg, 1.16 mmol) in MeOH (15 mL) at roomtemperature was added a solution of Pd/C 10% (50 mg. in MeOH ˜3 mL).Then the resulting solution was purged several times with H₂ with afinal pressure of 60 psi. The solution was stirred 4 h then filtratedthrough a Celite pad with a fritted glass funnel. The filtrate wasevaporated and the residue compound 37 was pure enough to use for thenext step without further purification.

¹H NMR: (300 MHz, acetone-d₆): δ 8.92 (broad s, 1H), 7.79-7.76 (m, 2H),7.60-7.47 (m, 3H), 7.12 (s, 4H), 3.32 (s, 1H), 2.81 (t, J=8 Hz, 2H),2.53 (t, J=8 Hz, 2H).

Step 2: N-Hydroxy-3-[4-(benzenesulfonylamino)-phenyl]-2-propanamide (38)

To a solution of 37 (1.16 mmol) in DMF (10 mL) at room temperature wereadded 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (EDC,266 mg, 1.39 mmol), and 1-Hydroxybenzotriazole hydrate (HOBT, 235 mg,1.74 mmol). The mixture was stirred 20 min. at room temperature thenNH₂OTHP (204 mg, 1.74 mmol) was added. The resulting mixture was heatedat 50° C. for 24 h then the DMF solvent was condensed under reducedpressure and the residue was dissolved in CH₂Cl₂, washed with asaturated aqueous solution of NaHCO₃. The combined organic extracts weredried over (MgSO₄) then evaporated. The crude compound was purified byflash chromatography using Hexane/acetone (7:3) as solvent mixture. Theresidue was then dissolved in MeOH (10 mL) then 10-camphorsulfonic acid(CSA, 135 mg, 0.58 mmol) was added. The mixture was stirred 2 h at roomtemperature then the solvents were evaporated under reduced pressure atroom temperature to avoid thermal decomposition. The crude was purifiedby flash chromatography using CH₂Cl₂/MeOH (9:1) as solvent mixturegiving the title compound 38 (237 mg, 64%, for the last 3 steps) as ayellow solid.

¹H NMR: (300 MHz, acetone-d₆): δ 8.91 (broad s, 1H), 7.78-7.76 (m, 2H),7.57-7.51 (m, 3H), 7.10 (broad s, 4H), 2.82 (broad s, 2H), 2.34 (broads, 2H), 1.07 (s, 1H), 0.85 (s, 1H).

Example 20 N-Hydroxy-4-[4-(benzenesulfonylamino)-phenyl]-butanamide (42)

Step 1: Methyl-4-(4-aminophenyl)-butanoate (40)

To a solution of 4-(4-aminophenyl)-butyric acid (5 g, 27.90 mmol) inMeOH (100 mL) at room temperature was added HCl conc. (37% 15 mL). Theresulting mixture was stirred overnight at 50° C. then treated with asaturated aqueous solution NaHCO₃ and Na₂CO₃ solid until pH 9. Thesolvent was evaporated under reduced pressure then the aqueous phase wasextracted several times with CH₂Cl₂. The crude material was purified byflash chromatography using CH₂Cl₂/MeOH as solvent mixture yielding 40(4.93 g, 91%) as orange solid.

¹H NMR: (300 MHz, acetone-d₆): δ 6.89 (d, J=8 Hz, 2H), 6.59 (d, J=8 Hz,2H), 4.40 (broad s, 1H), 3.60 (s, 3H), 2.48 (t, J=7 Hz, 2H), 2.28 (t,J=7 Hz, 2H), 1.82 (qt, J=7 Hz, 2H).

Step 2: 4-[4-(benzenesulfonylamino)-phenyl]-butyric acid (41)

To a solution of 40 (500 mg, 2.59 mmol) in CH₂Cl₂ at room temperaturewere added Et₃N (901 μμL, 6.48 mmol) followed by benzenesulfonylchloride (661 μL, 5.18 mmol). The mixture was stirred overnight at roomtemperature then treated with a saturated aqueous solution of NH₄Cl. Thephases were separated and the organic layer was extracted several timeswith CH₂Cl₂. The combined organic extracts were dried over (MgSO₄) thenevaporated under reduced pressure. The residue was dissolved in asolvent mixture of THF (25 mL) and water (25 mL) then LiOH (1.08 g, 25.9mmol) was added. The mixture was heated at 50° C. for 1 h then treatedwith HCl (1N) until pH2. The phases were separated and the aqueous layerwas extracted several times with AcOEt. The combined organic extractswere dried over (MgSO₄) then evaporated. The crude was purified by flashchromatography using CH₂Cl₂/MeOH (95:5) as solvent mixture yielding 41(800 mg, 96%) as a white solid

¹H NMR: (300 MHz, CDCl₃): δ 8.82 (1H, s broad), 7.77-7.74 (2H, m),7.55-50 (1H, m), 7.44-7.39 (2H, m), 7.05-6.97 (4H, m), 2.58 (2H, t, J=7Hz), 2.31 (2H, t, J=7 Hz), 2.17 (1H, s), 1.94-1.84 (2H, m).

Step 3: N-Hydroxy-4-[4-(benzenesulfonylamino)-phenyl]-butanamide (42)

To a solution 41 (800 mg, 2.59 mmol) in DMF (20 mL) at room temperaturewere added 1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride(EDC, 593 mg, 3.12 mmol), and 1-Hydroxybenzotriazole hydrate (HOBT, 524mg, 3.89 mmol). The mixture was stirred 20 min. at room temperature thenNH₂OTHP (455 mg, 3.89 mmol) was added. The resulting mixture was heatedat 50° C. for 24 h then the DMF solvent was evaporated under reducedpressure and the residue was dissolved in CH₂Cl₂, washed with asaturated aqueous solution of NaHCO₃. The combined organic extracts weredried over (MgSO₄) then evaporated. The crude compound was purified byflash chromatography using Hexane/acetone (7:3) as solvent mixture. Theresidue was then dissolved in MeOH (30 mL) then 10-camphorsulfonic acid(CSA, 300 mg, 1.30 mmol) was added. The mixture was stirred 2 h at 50°C. then the solvents were condensed under reduced pressure at roomtemperature to avoid thermal decomposition. The crude was purified byflash chromatography using CH₂Cl₂/MeOH (9:1) as solvent mixture givingthe title compound 42 (115 mg, 13%) as a yellowish solid.

¹H NMR: (300 MHz, CDCl₃): δ 7.79-7.76 (m, 2H), 7.61-7.48 (m, 3H),7.13-7.05 (m, 4H), 2.83 (broad s, 1H), 2.53 (t, J=7 Hz, 2H), 2.14-2.04(m, 2H), 1.83 (t, J=7 Hz, 2H).

Example 21 N-Hydroxy-4-(3-oxo-3-phenylpropenyl)-benzamide (45)

Step 1: 4-(3-oxo-3-phenylpropenyl)-benzoic acid (43)

Sodium methoxide (1.8 g, 33 3 mmol) was added to a stirred suspension of4-carboxybenzaldehyde (2.5 g, 16.6 mmol) and acetophenone (2.0 g uL, 166 mmol) in methanol (50 mL) at room temperature. The mixture was stirredat room temperature for 16 hours, and half of the volume of methanol wasremoved under reduced pressure. The mixture was poured into HCI 1M (50mL) (until pH=2) and ethyl acetate was added. The separated aqueouslayer was extracted with ethyl acetate (3×30 mL) dried (MgSO₄ anh.),filtered and evaporated. The residue was triturated withdichloromethane-hexanes (1:1) to afford 3 g of 43 (72% yield).

¹H NMR (300 MHz, CDCl₃); δ 7.50-7.87 (m, 7H), 8.04 (d, 2H, J=8 Hz), 8.16(d, 2H, J=8 Hz)

Step 2: 4-(3-oxo-3-phenylpropenyl)-N—(O-tetrahydropyranyl)-benzamide(44)

The carboxylic acid 43 (260 mg, 1.0 mmol) was dissolved in anhydrousCH₂Cl₂ (10 mL) and DCC (256 mg, 1.2 mmol) followed by NH₂OTHP (145 mg,1.2 mmol) were added. The mixture was allowed to stir at roomtemperature for 2 h. Added NH₄Cl sat. and extracted with EtOAc. Theorganic layer was dried over MgSO₄, filtered and the solvent wasevaporated under vacuum. (Purification by column chromatography using 1%MeOH/CH₂Cl₂ give the title compound which was used directly in the nextstep.

Step 3: N-Hydroxy-4-(3-oxo-3-phenylpropenyl)-benzamide (45)

The protected hydroxamic acid 44 (234 mg, 0.67 mmol) was dissolved inMeOH (7 mL) then CSA (31 mg, 0.13 mmol) was added. The mixture wasallowed to stir at reflux for 2 hours or until the reaction was completeby TLC. Added HCl 1N, extracted with EtOAc, dried the organic layer overanhydrous MgSO₄ and filtered. The solvent was evaporated under vacuum.Purification by column chromatography using 5% MeOH/CH₂Cl₂, gave thetitle compound.

¹H NMR (300 MHz, DMSO-d₆), δ 7.53-8.20 (m, 11H); 9.12 (br. s, 1H); 11.35(br. s, 1H)

Example 22 N-Hydroxy-4-(3-oxo-3-phenylpropyl)-benzamide (50)

Step 1: Methyl-4-(3-oxo-3-phenylpropenyl)-benzoate (46)

To 4-carbomethoxybenzaldehyde (79 mg, 0.48 mmol) and acetophenone (56μμL, 0.48 mmol) in anhydrous methanol (1.6 mL), was added neat sodiummethoxide (26 mg, 0.48 mmol). The mixture was stirred at roomtemperature overnight then heated to reflux for 1 hour, cooled down toroom temperature and added HCl 1N and EtOAc. The layers were separatedand the organic layer dried over anhydrous MgSO₄ and filtered. Thesolvent was evaporated under vacuum to afford a yellow solid, which wasrecrystallized from acetonitrile/water to give a pale yellow crystallinesolid.

¹H NMR (300 MHz, CDCl₃); δ 3.95 (s, 3H), 7.50-8.12 (m, 11H)

Step 2: Methyl-4-(3-oxo-3-phenylpropyl)-benzoate (47)

The aromatic enone 46 (321 mg, 1.20 mmol) was dissolved in anhydrous THF(6 mL) and anhydrous MeOH (6 ml). Added 2 small scoops of Pd 10% onactivated C, placed under an atmosphere of hydrogen and allowed to stirfor 2 hours at room temperature. Purged with nitrogen, filtered throughCelite and removed solvent by evaporation under vacuum. The benzylicalcohol is reoxidized to the ketone by the following procedure. Thecrude was taken back in anhydrous CH₂Cl₂ (10 mL), with 3 Å molecularsieves, TPAP (1 scoop) was added followed by NMO (212 mg, 1.8 mmol).Stirred at room temperature for 30 minutes and filtered through a plugof silica gel. Solvent was evaporated under vacuum and purified bycolumn chromatography using 10% EtOAc/Hexane.

¹H NMR (300 MHz, CDCl₃); δ 3.14 (t, 2H), 3.34 (t, 2H), 3.90 (s, 3H),7.30-7.60 (m, 6H), 7.92-7.99 (m, 4H).

Step 3: 4-(3-oxo-3-phenylpropyl)-benzoic acid (48)

To a solution of methyl ester 47 (195 mg, 0.73 mmol) in water/THF (1:1,0.07M) was added LiOH (46 mg, 1.1 mmol). The resulting solution wasstirred overnight at room temperature or until no starting material wasdetected by TLC. HCl 1N was added and the solution was extracted withEtOAc and the organic layer was dried over anhydrous MgSO₄. Filtrationand evaporation of the solvent under vacuum followed by purification bycolumn chromatography using 10% MeOH/CH₂Cl₂, gave the title compound.

¹H NMR (300 MHz, CDCl₃); δ 3.16 (t, 2H), 3.36 (t, 2H), 7.33-7.60 (m,5H), 7.93-8.06 (m, 4H).

Step 4: N-hydroxy-4-(3-oxo-3-phenylpropyl)-benzamide (50)

Following the procedure described in Example 21, Steps 2-3, butsubstituting compound 48 for carboxylic acid 4, the title compound wasobtained.

¹H NMR (300 MHz, DMSO-d₆); δ 2.97 (t, 2H), 3.38 (t, 2H), 7.34 (d, 2H,J=8 Hz), 7.45-7.70 (m, 5H), 7.96 (dd, 2H, J=8 Hz, 1 Hz), 11.14 (br. s,1H)

Example 23 N-Hydroxy-4-(3-oxo-3-phenyl-1-hydroxypropyl)-benzamide (53)

Step 1: 4-Carboxy-N—(O-tetrahydropyranyl)-benzamide (51)

Hydroxylamine-O-THP (3.9 g, 33 2 mmol) was added to a suspension of4-formylbenzoic acid (4.2 g, 27.7 mmol) and DCC (6.8 g, 33 2 mmol) indichloromethane (200 mL). The mixture was stirred at room temperatureovernight and quenched with saturated ammonium chloride. The separatedaqueous layer was extracted with ethyl acetate (3×100 ml) and thecombined organic layers were washed with brine, dried (MgSO₄ anh),filtered and evaporated. Flash chromatography of the residue (10%methanol in CH₂Cl₂), afforded (51).

¹H NMR (300 MHz, CDCl₃); δ ppm. 10.04 (s, 1H), 8.95 (s, 1H), 7.99 (d,2H, J=7.0 Hz), 7.93 (d, 2H, J=7.0 Hz), 5.1 (s, 1H), 3.60 (m, 2H), 1.60(m, 6H)

Step 2:4-(3-oxo-3-phenyl-1-hydroxypropyl)-N—(O-tetrahydropyranyl)-benzamide(52)

n-BuLi (1.4M/hexane, 1.6 mL, 2.2 mmol) was added to a 0° C. solution ofdiisopropylamine (337 μL, 2.4 mmol) in anhydrous THF (15 mL). Stirred at0° C. 10 minutes, then cooled to −78° C. Added acetophenone, thenstirred 30 minutes at −78° C. Cannulated into a −78° C. solution of thealdehyde 9 (50 mg, 2.0 mmol) in anhydrous THF (10 mL). Stirred 3 hoursat −78° C., then added NH₄Cl. Warmed to room temperature, extracted withEtOAc, dried over MgSO₄, filtered and evaporated solvent under vacuum.Purification by HPLC CH₃CN: H₂O: TFA 0.1%; 10-95% gave the titlecompound 52.

Step 3: N-Hydroxy-4-(3-oxo-3-phenyl-1-hydroxypropyl)-benzamide (53)

Following the same procedure as described in Example 21, Step 3, butsubstituting compound 52 for compound 44, the title compound wasobtained.

¹H NMR (300 MHz, DMSO-d₆); δ 3.20 (dd, 1H, J=4 Hz, J=16 Hz), 3.42 (dd,1H=16 Hz, 8 Hz), 5.20 (m, 1H), 7.44-8.18 (m, 9H), 11.15 (br. s, 1H),11.32 (br. s, 1H)

Example 24 N-Hydroxy-4-(3-phenylpropyl)-benzamide (56)

Step 1: 4-(3-phenylpropenyl)-benzoicacid/4-(3-phenyl-2-propenyl)-benzoic acid (54)

Allylbenzene (255 μμL, 1.9 mmol), 4-bromobenzoic acid (523 mg, 2 6mmol), Et₃N (0.91 mL, 6.5 mmol), Palladium (II) Acetate (16 mg, 0.052mmol), triphenylphosphine (60 mg, 0.21 mmol) and acetonitrile (5 mL)were stirred at reflux overnight in a round bottom flask. Added HCl 1N,extracted with EtOAc, dried the organic layer on anhydrous MgSO₄,filtered, evaporated solvent under vacuum. Purified by columnchromatography using 10% MeOH/CH₂Cl₂ yielded 90 mg (14%) of mixture oftwo regioisomers 54. The mixture was then submitted for hydrogenationwithout further characterization.

Step 2: 4-(3-phenylpropyl)-benzoic acid (55)

A mixture of regioisomeric olefins 54 (100 mg, 0.42 mmol) and Pd 10% onC (10 mg) in methanol (4 mL) was vigorously stirred under H₂ atmosphere(14 psi). The mixture was stirred for 2 hours at room temperature,filtered through Celite and evaporated to afford 55 as an oil. Flashchromatography of the residue gave 55 (88 mg, 88%).

¹H NMR (300 MHz, CDCl₃); δ ppm 8.10 (d, 2H, J=8.0 Hz), 7.35 (m, 7H),2.73 (m, 4H), 2.00 (m, 2H)

Step 3: N-Hydroxy-4-(3-phenylpropyl)-benzamide (56)

Following the same procedure as described in Example 21, Steps 2-3, butsubstituting compound 55 for compound 43, the title compound wasobtained as a beige solid. (24 mg, 26% yield)

¹H NMR (300 MHz, CD₃OD); δ (ppm) 7.63 (d, 2H, J=8.0 Hz); 7.38-7.05 (m,7H), 2.63 (m, 4H), 1.91 (m, 2H)

Example 25 N-Hydroxy-4-(4-phenylbutyl)-benzamide (61)

Step 1: 4-(1-butenyl-4-phenyl)-benzoicacid/4-(2-butenyl-4-phenyl)-benzoic acid (57/58)

Under nitrogen atmosphere in a 25 mL round bottomed flask were mixed:4-phenyl-1-butene (568 μL, 3.8 mmol), 4-bromobenzoic acid (634 mg, 3 2mmol), tris(dibenzylideneacetone)dipalladium(0) (87 mg, 0.1 mmol),tri-o-tolylphosphine (58 mg, 0.2 mmol), triethylamine (1.1 mL, 7.9 mmol)in N,N-dimethylformamide (7 mL, 0.5 M solution). The mixture was stirredfor 22 hours at 100° C. Then, the resulting suspension was cooled toroom temperature, filtered through Celite and rinsed with ethyl acetate.The filtrate was acidified with 1N HCl, the phases were separated andthe aqueous layer was extracted with ethyl acetate. The combined organiclayers were washed with water, brine, dried over MgSO₄, filtered andconcentrated. The resulting solid was triturated withhexane:dichloromethane (9:1) to give 367 mg (46%) of beige solid 57/58.

¹H NMR (300 MHz, (CD₃)₂CO): δ (ppm) 2.50-2.60 (m, 2H), 2.80 (t, 2H,J=9.0 Hz), 6.40-6.50 (m, 2H), 7.12-7.35 (m, 5H), 7.41 (d, 2H, J=9.0 Hz),7.92 (d, 2H, J=9.0 Hz).

Step 2: 4-(4-phenylbutyl)-benzoic acid (59)

Following the procedure described in Example 24, Step 2, butsubstituting compound 57/58 for compounds 54, the title compound wasobtained as a white solid in 92% yield.

¹H NMR (300 MHz, CD₃OD); δ (ppm) 1.60-1.75 (m, 4H), 2.65 (t, 2H, J=9.0Hz), 2.72 (t, 2H, J=9.0 Hz), 7.12-7.30 (m, 5H), 7.33 (d, 2H, J=9.0 Hz),7.96 (d, 2H, J=9.0 Hz)

Step 3: 4-(4-phenylbutyl)-N—(O-tetrahydropyranyl)-benzamide (60)

Under nitrogen atmosphere in a 25 mL round bottomed flask, to4-(4-phenylbutyl)benzoic acid 59 (341 mg, 1.3 mmol) in 5 mL ofN,N-dimethylformamide (0.3 M solution) was added the1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (308 mg, 1.6mmol) and the 1-hydroxybenzotriazole hydrate (272 mg, 2.0 mmol) at roomtemperature. The mixture was stirred for 30 minutes then, the2-(tetrahydropyranyl)hydroxylamine (235 mg, 2.0 mmol) was added and themixture was stirred for 4 days. The N,N-dimethylformamide was removedunder vacuum, the resulting oil was dissolved in ethyl acetate, washedwith water and brine, dried over MgSO₄, filtered and concentrated togive 95% yield of crude title compound 60.

¹H NMR (300 MHz, CD₃OD); δ (ppm) 1.50-1.75 (m, 10H), 2.65 (t, 2H, J=9.0Hz), 2.72 (t, 2H, J=9.0 Hz), 3.51 (d, 1H, J=15 Hz), 4.05 (t, 1H, J=15Hz), 5.05 (s, 1H), 7.10-7.35 (m, 7H), 7.75 (d, 2H, J=9.0 Hz), 10.60 (s,1H)

Step 4: N-Hydroxy-4-(4-phenylbutyl)-benzamide (61)

Under nitrogen atmosphere, to the crude oil in a 25 mL round bottomedflask, were added 5 mL of methyl alcohol (0.3 M solution) andcamphorsulfonic acid (333 mg, 1.4 mmol). The mixture was stirred for 2hours at room temperature. The methyl alcohol was removed under vacuumwithout heating and the resulting oil was purified by flashchromatography eluting methyl alcohol and dichloromethane (1:19). Thesolid was with hexane:dichloromethane (9:1) to give 212 mg (59%) ofbeige solid 61.

¹H NMR (300 MHz, (CD₃)₂CO): δ 1.66 (m, 4H), 2.65 (t, 2H, J=7.2 Hz), 2.70(t, 2H, J=7.1 Hz), 7.15-7.31 (m, 7H), 7.75 (d, 2H, J=7.8 Hz), 8.18(broad s, 1H), 10.68 (broad s, 1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 31.6 (t), 31.8 (t), 36.1 (t), 36.2 (t),2×126.4 (d), 127.8 (d), 2×129.1 (d), 2×129.2 (d), 2×129.3 (d), 130.6(s), 143.3 (s), 147.3 (s), 165.9 (s).

Example 26 N-Hydroxy-3-(3-phenylpropyl)-benzamide (64) Step 1:3-(3-phenylpropenyl)-benzoic acid (62)

Following the same procedure as described in Example 24, step 1, butsubstituting

4-bromobenzoic acid for 3-bromobenzoic acid, the title compound wasobtained as mixture of olefins. The mixture was submitted to the nextstep without purification.

¹H NMR (300 MHz, CDCl₃); δ (ppm); 3.6 (dd, 2H, CH₂); 6.4 (dd, 2H,vinylic); 7.0-7.5 (m, 8H, CHAr); 8.0 (s, 1H, CHAr)

Step 2: 3-(3-phenylpropyl)-benzoic acid (63)

Following the same procedure as described in Example 24, Step 2, butsubstituting compound 62 for compound 54, the title compound wasobtained in 52% yield and submitted to the next step without furtherpurification.

¹H NMR (300 MHz, CDCl₃); δ (ppm); 2.0 (m, 2H, CH₂); 2.7 (m, 4H, 2CH₂);7.0-7.4 (m, 8H, CHAr); 8.0 (s, 1H, CHAr)

Step 3: N-Hydroxy-3-(3-phenylpropyl)-benzamide (64)

Following the procedure described in Example 25, Step 3-4, butsubstituting compound 63 for compound 59, the title compound wasobtained. Purification by flash chromatography using CH₂Cl₂: MeOH(9.5:0.5) gave compound 64 in 20% yield.

¹H NMR (300 MHz, DMSO-d₆); δ 1.8 (m, 2H, CH₂); 2.8 (m, 4H, CH₂); 7.0-7.4(m, 7H, CHAr); 7.6 (s, CHAr); 9.0 (s, NH); 11.2 (s, OH)

Example 27 N-Hydroxy-3-(2-phenylethyl)-benzamide (68)

Step 1: 3-(2-phenylethenyl)-benzoic acid (65/66)

A 1.0 M solution of lithium bis(trimethylsilyl) amide (3.3 mL, 3.3 mmol)in THF was added to a stirred suspension of benzyltriphenylphosphoniumbromide (1.44 g, 3.6 mmol) in THF (35 mL) at 0° C. The resulting orangesolution was added via cannula to a mixture of 3-carboxybenzaldehyde(500 mg, 3.3 mmol) and lithium bis(trimethylsilyl)amide (3.3 mL, 3.3mmol) in THF (10 mL). The mixture was stirred overnight at roomtemperature. A 1N solution of HCl (75 mL) and ethyl acetate (75 mL) wereadded and the separated aqueous layer was extracted with ethyl acetate(3×50 mL), dried (MgSO₄ anh.) filtered and evaporated. The residue waspurified by HPLC (10:95 CH₃CN: H₂O, TFA 0.1%) to afford 130 mg of thetitle compound (17%)

¹H NMR (300 MHz, CDCl₃); δ (ppm) (1:1) E:Z mixture 8.22 (s, 1H), 7.98(s, 1H), 7.90-7.10 (m, 16H), 6.70 (d, 1H, J=15.0 Hz), 6.62 (d, 1H,J=15.0 Hz)

Step 2: 3-(2-phenylethyl)-benzoic acid (67)

Following the same procedure as described in Example 24, Step 2, butsubstituting compounds 65/66 for compound 54, the title compound wasobtained quantitatively.

¹H NMR (300 MHz, CDCl₃); δ (ppm) 2.98 (m, 4H); 7.30 (m, 7H); 7.99 (m,2H)

Step 3: N-Hydroxy-3-(2-phenylethyl)-benzamide (68)

Following the same procedure as described in Example 25, Step 3 and 4,but substituting compound 67 for compound 59, the title compound wasobtained in 22% yield.

¹H NMR (300 MHz, DMSO-d₆); δ (ppm) 2.82 (s, 4H); 7.03-7.08 (m, 8H); 7.62(s, 1H); 8.98 (br. s, 1H); 11.15 (br. s, 1H)

Example 28 N-Hydroxy-4-(2-thiophenyl)-ethyl benzamide (70)

Step 1: 4-(2-thiophenyl)-ethyl benzoic acid (69)

According to the published procedure (Gareau et al., Tet. Lett., 1994,1837), under nitrogen atmosphere in a 50 mL round bottomed flaskcontaining 4-vinylbenzoic acid (1.0 g, 6.75 mmoles) in 10 mL of benzene(0.7 M) was added benzenethiol (797 μL, 7.76 mmoles) followed by VAZO™(Aldrich Chemical Company, 495 mg, 2.02 mmoles). The mixture was stirredfor 12 hours at reflux. The resulting solution was cooled at roomtemperature and the solvent was evaporated under vacuo. The solid waspurified by trituration using hexane and dichloromethane to afford 1.94g (85%) of white solid.

¹H NMR (300 MHz, CDCl₃): δ 3.01 (t, 2H, J=8.4 Hz), 3.28 (dd, 2H, J=7.2,7.8 Hz), 7.21 (tt, 1H, J=1.2, 7.2 Hz), 7.34 (t, 2H, J=8.1 Hz), 7.38-7.43(m, 1H), 7.41 (d, 2H, J=8.4 Hz), 7.97 (d, 2H, J=8.1 Hz).

Step 2: N-Hydroxy-4-(2-thiophenyl)-ethyl benzamide (70)

Under nitrogen atmosphere in a 50 mL round bottomed flask containing4-(2-thiophenyl)-ethyl benzoic acid (600 mg, 2.32 mmoles) in 12 mL ofN,N-dimethylformamide (0.2 M) was added1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (579 mg,3.02 mmoles) and 1-hydroxybenzotriazole hydrate (377 mg, 2.79 mmoles) atroom temperature. The mixture was stirred 30 minutes then, hydroxylaminehydrochloride (242 mg, 3.48 mmoles) and triethylamine (971 μL, 6.97mmoles) was added and the mixture was stirred for 12 hours at 50° C. TheN,N-dimethylformamide was removed under vacuo and the resulting oil wasdissolved in ethyl acetate, washed with water, saturated sodium hydrogencarbonate solution, water and brine. The organic layer was dried overanhydrous magnesium sulfate, filtered and concentrated under vacuo. Thecrude solid was purified by trituration using hexane and dichloromethaneto afford 450 mg (71%) of a beige solid.

RP-HPLC (Hewlett-Packard 1100, column C18 HP 4.6×250 mm, flow 1 mL/min,10-95% CH₃CN/H₂O in 42 min with 0.1% TFA); Purity: 95.8% (220 nm), 93.2%(254 nm).

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 2.98 (t, 2H, J=7.2 Hz), 3.26 (dd, 2H,J=6.6, 8.4 Hz), 7.21 (tt, 1H, J=1.5, 6.9 Hz), 7.31-7.42 (m, 6H), 7.77(d, 2H, J=9.3 Hz), 8.08 (broad s, 1H), 10.69 (broad s, 1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 34.8 (t), 35.9 (t), 126.7 (d), 127.9(d), 2×129.6 (d), 2×129.7 (d), 2×129.9 (d), 131.3 (s), 137.3 (s), 145.0(s).

Elemental Analysis; Calc for C₁₅H₁₅O₂NS×0.1H₂O: % C=75.31, % H=7.14, %N=5.17. Found: % C=75.2±0.1, % H=7.41±0.07, % N=5.17±0.01.

N-Hydroxy-4-(2-benzenesulfonyl)-ethyl benzamide (73) Step 1:4-(2-benzenesulfonyl)-ethyl benzoic acid (72)

Under nitrogen atmosphere in a 100 mL round bottomed flask containing4-(2-thiophenyl)-ethyl benzoic acid (69) (600 mg, 2.32 mmoles) in 20 mLof dichloromethane (0.1 M) at 0° C. was added portionwise3-chloroperbenzoic acid (Aldrich Chemical Co., 57-86% pure solid by, 2g, 6.97 mmoles), as described by Nicolaou et al., J. Am. Chem. Soc.,114: 8897 (1992). The mixture was allowed to reach room temperature andwas stirred for 1 hour. Dimethyl sulfide (5 mL) was added, the mixturewas diluted in dichloromethane and washed 3 times with water. Theorganic layer was dried over anhydrous magnesium sulfate, filtered andthe solvent were evaporated in vacuo to afford 3 g of white solid. Thismixture of 3-chlorobenzoic acid and the desired4-(2-benzenesulfonyl)-ethyl benzoic acid was placed in a 125 mLErlenmeyer flask, dissolved in 30 mL of dichloromethane and treated withan excess of freshly prepared diazomethane solution in diethyl ether(0.35 M). Nitrogen was bubbled to removed the excess of diazomethane andsolvents were evaporated under vacuum. The resulting solid was purifiedby flash chromatography, eluting with 20% ethyl acetate: 80% hexane toafford 341.6 mg (48%) of the corresponding ester. Saponification of thisester was done using the same procedure as described in Example 1, step2, to afford 312.4 mg (96%) of 4-(2-benzenesulfonyl)-ethyl benzoic acid(72)

¹H NMR (300 MHz, CDCl₃): δ 3.06-3.11 (m, 2H), 3.56-3.61 (m, 2H), 7.37(d, 2H, J=8.4 Hz), 7.67 (tt, 2H, J=1.5, 7.2 Hz), 7.76 (tt, 1H, J=1.2,7.5 Hz), 7.93 (d, 2H, J=8.7 Hz), 7.97 (dd, 2H, J=1.8, 6.9 Hz).

Step 2: N-Hydroxy-4-(2-benzenesulfonyl)-ethyl benzamide (73)

Following the procedure described for N-hydroxy-4-(2-thiophenyl)-ethylbenzamide, but substituting 4-(2-benzenesulfonyl)-ethyl benzoic acid for4-(2-thiophenyl)-ethyl benzoic acid, the title compound was obtained asa beige solid.

RP-HPLC (Hewlett-Packard 1100, column C18 HP 4.6×250 mm, flow 1 mL/min,10-95% CH₃CN/H₂O in 42 min with 0.1% TFA); Purity: 98.8% (220 nm), 97.6%(254 nm).

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 2.98 (t, 2H, J=7.2 Hz), 3.26 (dd, 2H,J=6.6, 8.4 Hz), 7.21 (tt, 1H, J=1.5, 6.9 Hz), 7.31-7.42 (m, 6H), 7.77(d, 2H, J=9.3 Hz), 8.08 (broad s, 1H), 10.69 (broad s, 1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 25.2 (t), 34.3 (t), 55.6 (t), 128.0(d), 2×128.8 (d), 129.4 (d), 2×130.2 (d), 131.1 (s), 134.5 (d), 140.7(s), 145.5 (s), 165.8 (s).

N-Hydroxy-4-(2-benzenesulfoxide)-ethyl benzamide (71)

According to the procedure described by Van Der Borght et al., J. Org.Chem., 65: 288 (2000), under anitrogen atmosphere in a 10 mL roundbottomed flask containing N-hydroxy-4-(2-thiophenyl)-ethyl benzamide(70) (50 mg, 0.18 mmol) in 2 mL of methanol (0.1 M) was added telluriumdioxide (3 mg, 0.018 mmol) followed by a solution 35% in water ofhydrogen peroxide (32 μL, 0.36 mmol). The mixture was stirred for fivedays and then brine was added. The aqueous layer was extracted 3 timeswith ethyl acetate and the combined organic layers were dried overanhydrous magnesium sulfate, filtered and the solvent were evaporatedunder vacuo. The resulting solid (43.3 mg) was purified by triturationusing acetonitrile to afford 10 mg (20%) of beige solid.

RP-HPLC (Hewlett-Packard 1100, column C18 HP 4.6×250 mm, flow 1 mL/min,10-95% CH₃CN/H₂O in 42 min with 0.1% TFA); Purity: 98.8% (220 nm), 97.9%(254 nm).

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 2.76-2.91 (m, 1H), 3.00-3.29 (m, 3H),7.34 (d, 2H, J=8.4 Hz), 7.55-7.62 (m, 3H), 7.70 (dd, 2H, J=1.5, 8.1 Hz),7.76 (d, 2H, J=8.1 Hz), 8.08 (broad s, 1H), 10.70 (broad s, 1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 28.3 (t), 57.8 (t), 2×124.8 (d), 128.0(d), 2×129.6 (d), 2×130.0 (d), 131.5 (d), 144.1 (s), 145.7 (s).

Example 29 N-Hydroxy-3-[4-(3-phenylpropyl)-phenyl]-propanamide (77)

Step 1: 3-(4-bromophenyl)-propanoic acid (74)

Under nitrogen atmosphere in a 250 mL round bottomed flask containing4-bromocinnamic acid (5.0 g, 22 mmoles) in 45 mL ofN,N-dimethylformamide (0.5 M) was added benzenesulfonylhydrazide (7.6 g,44 mmoles). The mixture was stirred at reflux for 12 hours. The solutionwas cooled at room temperature, aqueous saturated ammonium chloride wasadded and the aqueous layer was extracted with ethyl acetate 3 times.Combined organic layers were washed with water and brine, dried overanhydrous magnesium sulfate, filtered and concentrated under vacuo. Theresulting solid was purified by flash chromatography eluting with 5%methanol:95% dichloromethane to afford 3.66 g (73%) of beige solid.

¹H NMR (300 MHz, CDCl₃): δ 2.66 (t, 2H, J=7.5 Hz), 2.91 (d, 2H, J=7.5Hz), 7.08 (d, 2H, J=8.4 Hz), 7.41 (d, 2H, J=8.4 Hz).

Step 2: N-Hydroxy-3-(4-bromophenyl)-propanamide (75)

Following a procedure analogous to that described for the preparation of70, 1.54 g (39%) of the title compound was obtained.

¹H NMR (300 MHz, CDCl₃): δ 2.39 (t, 2H, J=7.8 Hz), 2.89 (d, 2H, J=7.2Hz), 7.18 (d, 2H, J=8.1 Hz), 7.42 (d, 2H, J=8.7 Hz), 8.18 (broad s, 1H),9.98 (broad s, 1H).

Step 3: N-Hydroxy-3-[4-(3-phenyl-1-propenyl)-phenyl]-propanamide andN-Hydroxy-3-[4-(3-phenyl-2-propenyl)-phenyl]-propanamide (76)

Following a procedure analogous to that described in Example 25, step 1,but substituting N-hydroxy-3-(4-bromophenyl)-propanamide (75) (250 mg,1.02 mmol) for 4-bromobenzoic acid and allyl benzene (163 μL, 1.2 mmol)for 4-phenyl-1-butene, to yield 155.4 mg (54%) of the mixed titlecompounds.

¹H NMR (300 MHz, CDCl₃): δ 2.39 (m, 2H), 2.88 (t, 2H, J=8.4 Hz), 3.51(t, 2H, J=8.1 Hz), 6.32-6.53 (m, 2H), 7.14-7.44 (m, 9H), 8.60 (broad s,1H), 10.04 (broad s, 1H).

Step 4: N-Hydroxy-3-[4-(3-phenylpropyl)-phenyl]-propanamide (77)

Following a procedure analogous to that described in Example 24, step 2,but substituting the mixture ofN-hydroxy-3-[4-(3-phenyl-1-propenyl)-phenyl]-propanamide andN-hydroxy-3-[4-(3-phenyl-2-propenyl)-phenyl]-propanamide (155 mg, 0.55mmol) for olefins 54, 155.4 mg (99%) of the title compound was obtained.

RP-HPLC: (Hewlett-Packard 1100, column C18 HP 4.6×250 mm, flow 1 mL/min,10-95% CH₃CN/H₂O in 42 min with 0.1% TFA); Purity: 99.9% (220 nm) (2peaks but same compound proven by LCMS, 99.9% (254 nm). ¹H NMR (300.072MHz, (CD₃)₂CO): δ 1.91 (quintuplet, 2H, J=8.1 Hz), 2.38 (t, 2H, J=7.8Hz), 2.61 (q, 4H, J=9.6 Hz), 2.87 (t, 2H, J=7.2 Hz), 7.12-7.29 (m, 9H),8.42 (broad s, 1H), 10.01 (broad s, 1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 26.3 (t), 28.7 (t), 29.8 (t), 30.3 (t),30.7 (t), 121.1 (d), 3×123.7 (d), 3×123.8 (d), 133.9 (s), 133.4 (s),137.8 (s), 164.9 (s).

Elemental Analysis; Calc for C₁₈H₂₁O₂N×0.1H₂O: % C=75.81, % H=7.49, %N=4.91. Found: % C=75.7±0.3, % H=7.54±0.02, % N=4.85±0.03.

Example 30

Step 1: Ethyl 3-(4-nitrophenyl),2-isopropyl propanoate (78)

To a precooled solution of diisopropylamine (34 7 mmol) in THF (30 mL)under nitrogen was added dropwise a 1.0 M solution of n-butyllithium(33.3 mmol). The resulting light yellow solution was stirred at −78° C.over 30 minutes and transferred via canula to a precooled (−78° C.)solution of ethyl isovalerate (34.7 mmol) in THF (50 mL). The mixturewas stirred at −78° C. over 1 hour and a 4-nitrobenzyl bromide (13.9mmol) solution in THF (20 mL) at room temperature was transferreddropwise via canula to the enolate solution which turned deep red. Themixture was stirred over 15 minutes and the reaction was quenched withaqueous saturated ammonium chloride solution (NH₄Cl). The mixture wasallowed to warm to room temperature over 1 hour and turned brown uponwarming. It was poured into a large volume of saturated NH₄Cl solutionand the layers were separated. The aqueous layer was extracted twicewith diethyl ether and the combined organic layers were washed withbrine, dried over magnesium sulfate and concentrated in vacuo. Theresidue was purified by flash chromatography on silica gel using ethylacetate and hexanes (10:90) as the eluent, yielding 73% of the puretitle compound 78 as a light yellow oil.

Step 2: Ethyl 3-(4-aminophenyl),2-isopropyl propanoate (79):

To a hydrogen flushed (vacuum/H₂, 3 times) solution of 1 (1.88 mmol) inmethanol (10 mL) was added 10% palladium on charcoal (0.018 mmol)previously quenched with methanol in a separate flask. The blackheterogeneous resulting mixture was stirred at room temperature underhydrogen atmosphere (1 atm) over 20 hours. The hydrogen was thenevacuated by vacuum and replaced with air. Then, the mixture wasfiltered through celite, rinsing with methanol while making sure the padnever gets dry. The filtrate was concentrated to a red oil. The residuewas purified by flash chromatography on silica gel using ethyl acetateand hexanes (30:70) as the eluent, yielding 73% of the pure titlecompound 79 as a light red oil.

Steps 3-5: (81)

Compound 79 was coupled with benzenesulfonyl chloride in the presence oftriethylamine according to the procedure described in Example 1, step 1,to afford the sulfonamide 80. Ester hydrolysis and coupling withhydroxylamine were then accomplished as described in Example 28 toafford the hydroxamic acid 81.

¹H NMR: (Acetone-d₆) δ (ppm): 9.76 (bs, 1H), 8.83 (bs, 1H), 7.74 (d,J=8.2 Hz, 2H), 7.59-7.49 (m, 3H), 7.04 (s, 4H), 2.83-2.73 (m, 3H), 1.83(sext, J=6.9 Hz, 1H), 1.00 (d, J=6.9 Hz, 3H), 0.93 (d, J=6.9 Hz, 3H).

HRMS: 344.1195 (M⁺-H₂O) (calc.); 344.1200±0.0010 (found).

Example 31

Compound 82 was obtained in good yield from commercially availablebromoaminopyridine through a palladium catalyzed coupling withtert-butyl acrylate. Treatment of 82 with 4-phenylbenzenesulfonylchloride afforded a mixture of sulfonamide 84 and bis-sulfonamide 83,which was converted to 84 upon chromatographic isolation followed bybasic methanolysis. Acidic cleavage of the t-butyl ester was effected bytreatment of 84 with aqueous formic acid and a tert-butyl cationscavenger to afford the acrylic acid 85 in quantitative yield. Finally,coupling of 85 with o-phenylenediamine in the presence ofbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(BOP) afforded the anilide 86.

Data for 86:

¹H NMR: (300.07 MHz; CD₃OD): δ (ppm): 8.23 (d, J=1.9, 1H); 8.03 (bd,J=8.5; 2H); 7.96 (dd, J=1.9, 9.1; 1H); 7.76 (bd, J=8.5, 2H); 7.63 (dd,J=1.4, 8.2); 7.53 (J=15.5; 1H), 7.48-7.36 (m, 3H); 7.29 (d, J=9.1, 1H)7.18 (dd, J=1.4, 8.0, 1H); 7.03 (dt, J=1.4, 7.8, 1H); 6.86 (d, J=1.4,7.9, 1H) 6.76 (d, J=15.6, 1H) 6.75-6.69 (m, 1H); 4.85 (bs, 4H).

¹³C NMR: (75.5 MHz; CD₃OD) δ (ppm): 166.4; 154.7; 146.9; 146.2; 143.1;141.1; 140.6; 138.6; 137.9; 130.1; 129.5; 128.8; 128.5; 128.3; 126.7;125.6; 125.0; 122.1; 120.8; 119.5; 118.6; 114.9

MS: calc for C₂₆H₂₂O₃N₄S: 470.556. found: 471.5 for [M+H] (lowresolution MS).

By procedures analogous to those described in Examples 1-31 above, thefollowing compounds were synthesized:

¹H NMR: (300 MHz, CD₃OD): δ=7.76-7.74 (1H, m), 7.58-7.48 (4H, m), 7.22(2H, d, J=7.5 Hz), 7.10 (1H, t, J=5.1 Hz), 6.41 (1H, d broad, J=14.7Hz).

¹H NMR: (300 MHz, CD₃OD): δ=7.79 (2H, d, J=8.1 Hz), 7.56-7.46 (5H, m),7.17 (2H, d, J=8.1 Hz), 6.39 (1H, d, J=15.9 Hz).

Analysis: C₁₅H₁₃N₂O₄SCl×0.1H₂O, ×0.3 TFA Found: C=48.26%, H=3.58%,N=6.97%, S=7.86%. Calc.: C=48.19%, H=3.50%, N=7.20%, S=8.25%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.85 (1H, s br), 10.70 (1H, s br), 8.99(1H, s), 8.37 (2H, d, J=9 Hz), 8.01 (2H, d, J=9 Hz), 7.44 (2H, d, J=8.7Hz), 7.33 (1H, d, J=15.3 Hz), 7.12 (2H, d, J=8.4 Hz), 6.31 (1H, d,J=15.9 Hz).

Analysis: C₁₅H₁₃N₃O₆S×0.4H₂O, ×0.3 TFA Found: C=46.39%, H=3.49%,N=10.44%, S=7.92%. Calc.: C=46.29%, H=3.51%, N=10.38%, S=7.92%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.70 (1H, s br), 10.33 (1H, s br), 8.99(1H, s br), 7.44-7.26 (5H, m), 7.12 (2H, d, J=8.7 Hz), 7.06 (1H, d,J=8.4 Hz), 6.30 (1H, d, J=16.2 Hz), 3.78 (3H, s), 3.75 (3H, s)

Analysis: C₁₇H₁₈N₂O₆S×0.2H₂O Found: C=53.56%, H=5.03%, N=7.71%, S=8.01%.Calc.: C=53.45%, H=4.86%, N=7.33%, S=8.39%.

¹H NMR: (CD₃OD) δ (ppm): 7.78 (d, J=7.1 Hz, 1H), 7.56-7.45 (m, 3H), 7.24(d, J=8.5 Hz, 2H), 7.12 (d, J=8.8 Hz, 2H), 7.06 (s, 1H), 2.00 (d, J=1.4Hz, 3H).

¹³C NMR: (CD₃OD) δ (ppm): 135.2, 132.9, 128.1, 127.7, 125.5, 124.6,124.1, 122.3, 116.8, 115.6, 8.4.

¹H NMR: (Acetone-d₆) δ (ppm): 9.86 (bs, 1H), 8.86 (bs, 1H), 7.83 (bs,1H), 7.76 (d, J=6.7 Hz, 1H), 7.62-7.48 (m, 3H), 7.10-7.03 (m, 4H),2.87-2.79 (m, 3H), 2.56-2.39 (m, 2H), 1.05 (d, J=6.6 Hz, 3H).

HRMS: 334.0987 (calc.); 334.0991±0.0010 (found).

¹H NMR: (300 MHz, DMSO d₆): δ=10.94 (1H, s broad), 10.65 (1H, s broad),8.95 (1H, s Broad), 8.73-8.71 (1H, m), 8.24-8.21 (2H, m), 8.05 (1H, m),7.74-7.63 (3H, m), 7.33-7.23 (2H, m), 7.06-7.04 (2H, m), 6.24 (1H, d,J=15.3).

Analysis: C₁₉H₁₆N₂O₄S×0.5H₂O Found: C=60.31%, H=4.58%, N=7.43%. Calc.:C=60.46%, H=4.54%, N=7.42%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.65 (2H, s broad), 8.48 (1H, s),8.15-8.08 (2H, m), 8.00 (1H, d, J=7.5 Hz), 7.77 (1H, d, J=9 Hz),7.70-7.62 (2H, m), 7.39 (2H, d, J=8.4 Hz), 7.28 (1H, d, J=15.6 Hz), 7.15(2H, d, J=8.4 Hz), 6.26 (1H, d, J=15.6 Hz).

Analysis: C₁₉H₁₆N₂O₄S×0.2H₂O, ×0.5 TFA Found: C=56.01%, H=3.94%,N=6.60%, S=7.41%. Calc.: C=55.99%, H=3.97%, N=6.53%, S=7.47%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.91 (1H, s), 10.69 (1H, s br), 8.06-7.98(3H, m), 7.57-7.46 (4H, m), 7.34 (1H, d, J=15.9 Hz), 7.21 (2H, d, J=8.4Hz), 6.33 (1H, d, J=15.9 Hz).

¹H NMR: (300 MHz, DMSO d₆): δ=8.69-8.8 (1H, m), 8.02-8.01 (2H, m),7.61-7.59 (1H, m), 7.52-7.43 (3H, m), 7.25 (2H, d, J=7.5 Hz), 6.37 (1H,d, J=15.9 Hz).

Analysis: C₁₄H₁₃N₃O₄S×0.9 TFA Found: C=45.36%, H=3.51%, N=9.77%,S=7.09%. Calc.: C=44.97%, H=3.32%, N=9.96%, S=7.60%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.91 (1H, s), 10.62 (1H, s br), 8.45 (1H,8.1 Hz), 8.36 (1H, d, J=8.7 Hz), 8.25 (1H, d, J=6.9 Hz), 7.65-7.59 (2H,m), 7.37-7.34 (2H, m), 7.29-7.23 (2H, m), 7.06 (2H, d, J=8.7 Hz), 6.25(1H, d, J=15.9 Hz) 2.80 (6H, s).

¹H NMR: (300 MHz, DMSO d₆): δ=10.82 (1H, s br), 9.95 (1H, s br), 9.12(1H, s br), 7.70 (4H, s), 7.46 (1H, d, J=15.9 Hz), 6.79 (1H, d, J=8.7Hz), 6.68 (1H, s), 6.56-6.51 (2H, m), 3.65 (3H, s), 3.62 (3H, s).

¹H NMR: (300 MHz, DMSO d₆): δ=10.63 (1H, s), 10.36 (1H, s br), 9.13-9.12(1H, m), 8.93 (1H, s br), 8.51 (1H, d, J=8.1 Hz), 8.40 (1H, d, J=7.2Hz), 8.28 (1H, d, J=8.4 Hz), 7.75-7.70 (2H, m), 7.30-720 (3H, m), 7.09(2H, d, J=8.4 Hz) 6.21 (1H, d, J=15.9 Hz).

Analysis: C₁₈H₁₅N₃O₄S×1.1H₂O Found: C=55.72%, H=4.45%, N=10.64%,S=6.93%. Calc.: C=55.55%, H=4.45%, N=10.80%, S=8.24%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.72 (1H, s br), 10.07 (1H, s), 7.53-7.51(2H, m), 7.43-7.34 (4H, m), 7.26-7.19 (4H, m), 6.38 (1H, d, J=15.6 Hz),4.51 (2H, s).

Analysis: C₁₆H₁₆N₂O₄S×0.4 TFA Found: C=53.60%, H=4.46%, N=7.36%,S=7.81%. Calc.: C=53.38%, H=4.37%, N=7.41%,0=20.32%, S=8.48%, F=6.03%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.63 (1H, s br), 10.56 (1H, s), 8.67 (1H,s), 8.29 (1H, d, J=6.9 Hz), 7.89-7.85 (2H, m), 7.75 (1H, d, J=8.4 Hz),7.59 (1H, t, J=7.2 Hz), 7.47-7.38 (3H, m), 7.27 (1H, d, J=15.6 Hz), 7.15(2H, d, J=8.7 Hz), 6.25 (1H, d, J=15.9 Hz).

¹H NMR: (300 MHz, DMSO d₆): δ=10.72 (2H, s), 8.98 (1H, s br), 7.97 (4H,s), 7.55 (2H, s), 7.45 (2H, d, J=8.7 Hz), 7.33 (1H, d, J=15.9 Hz), 7.13(2H, d, J=8.7 Hz), 6.32 (1H, d, J=15.9 Hz).

1H NMR: (300 MHz, DMSO d₆): δ=10.75 (2H, m), 7.65-7.64 (1H, m),7.53-7.45 (4H, m), 7.35 (1H, d, J=16.2 Hz), 7.29 (1H, d, J=3.9 Hz), 7.20(2H, d, J=8.7 Hz), 7.12 (1H, t, J=3.6 Hz), 6.34 (1H, d, J=15.6 Hz).

Analysis: C₁₇H₁₄N₂O₄S₃×0.1H₂O, ×1.0 TFA Found: C=43.83%, H=3.26%,N=5.73%, S=18.15%. Calc.: C=43.69%, H=2.93%, N=5.36%, S=18.42%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.72 (1H, s), 8.91 (1H, d, J=1.8 Hz),8.80-8.78 (1H, m), 8.13 (1H, d, J=7.8 Hz), 7.63-7.59 (1H, m), 7.46 (2H,d, J=8.7 Hz), 7.33 (1H, d, J=15.6 Hz), 7.14 (2H, d, J=8.7 Hz), 6.32 (1H,d, J=15.9 Hz).

¹H NMR: (300 MHz, DMSO d₆): δ=10.54 (1H, s), 7.73 (2H, d, J=8.4 Hz),7.58 (2H, d, 8.4 Hz), 7.43 (2H, d, J=8.4 Hz), 7.32 (1H, d, J=15.6 Hz),7.15 (2H, d, J=8.4 Hz), 6.30 (1H, d, J=15.9 Hz), 1.25 (9H, s).

Analysis: C₁₉H₂₂N₂O₄S×0.3H₂O, 0.6 TFA Found: C=54.17%, H=5.25%, N=6.32%,S=6.85%. Calc.: C=54.12%, H=5.22%, N=6.25%, S=7.15%.

¹H NMR: (300 MHz, DMSO d₆): δ=11.02 (1H, s), 10.70 (1H, s), 8.99 (1H, sbr), 8.03 (1H, d, J=1.8 Hz), 7.76-7.67 (2H, m), 7.45 (2H, d, J=8.1 Hz),7.33 (1H, d, J=15.6 Hz), 7.13 (2H, d, J=8.4 Hz), 6.31 (1H, d, J=16.2Hz).

Analysis: C₁₅H₁₂N₂O₄SCl₂×0.3H₂O Found: C=45.96%, H=3.11%, N=7.21%,S=8.06%. Calc.: C=45.89%, H=3.23%, N=7.13%, S=8.17%.

¹H NMR: (300 MHz, Acetone d₆): δ=8.81 (1H, d, J=8.4 Hz), 8.34 (2H, d,J=7.2 Hz), 8.20 (1H, d, J=8.1 Hz), 8.05 (1H, d, J=7.5 Hz), 7.75-7.59(4H, m), 7.53-7.41 (4H, m), 7.23-7.07 (4H, m), 6.89-6.86 (2H, m), 6.75(1H, d, J=15.3 Hz).

Analysis: C₂₅H₂₁N₃O₃S×0.4H₂O, 0.6 TFA Found: C=60.68%, H=4.36%, N=8.11%,S=6.15%. Calc.: C=60.62%, H=4.35%, N=8.09%, S=6.18%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.7 (1H, s br), 10.45 (1H, s br), 8.96(1H, s br), 7.64 (2H, d, J=8.1 Hz), 7.38 (2H, d, J=8.4 Hz), 7.32-7.29(3H, m), 7.09 (2H, d, J=8.4 Hz), 6.29 (1H, d, J=16.2 Hz), 2.30 (3H, s).

Analysis: C₁₆H₁₆N₂O₄S×1.6H₂O, ×1.6 TFA Found: C=42.26%, H=3.62%,N=5.45%, S=6.09%. Calc.: C=42.42%, H=3.86%, N=5.15%, S=5.9%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.71 (1H, s), 10.67 (1H, s), 9.00 (1H, sbr), 7.96 (1H, d, J=2.4 Hz), 7.85 (1H, d, J=8.4 Hz), 7.69 (1H, dd, J=8.4Hz and 2.1 Hz), 7.47 (2H, d, J=8.4 Hz) 7.35 (1H, d, J=15.9 Hz), 7.13(2H, d, J=8.7 Hz), 6.33 (1H, d, J=15.9 Hz).

Analysis: C₁₅H₁₂N₂O₄SCl₂×0.3H₂O, ×0.3 AcOEt Found: C=46.30%, H=3.27%,N=6.56%, S=7.57%. Calc.: C=46.43%, H=3.61%, N=6.68%, S=7.65%.

¹H NMR: (300 MHz, DMSO d₆): δ=1.65 (1H, s br), 10.45 (1H, s br), 8.96(1H, s br), 7.42 (2H, d, J=8.1 Hz), 7.31 (1H, d, J=15.6 Hz), 7.22 (2H,s), 7.01 (2H, d, J=8.1 Hz), 6.30 (1H, d, J=15.9 Hz), 4.24-4.16 (2H, m),2.93-2.84 (1H, m), 1.18-1.14 (18H, m).

Analysis: C₂₄H₃₂N₂O₄S×1.10H₂O Found: C=62.14%, H=7.17%, N=6.20%,S=6.71%. Calc.: C=62.07%, H=7.42%, N=6.03%, S=6.9%.

¹H NMR: (300 MHz, DMSO d₆): δ=11.18 (1H, s br), 10.69 (2H, m), 7.83-7.82(1H, m), 7.68 (1H, m), 7.43 (2H, d, J=8.1 Hz), 7.32 (1H, d, J=15.3 Hz),7.13 (2H, d, J=8.1 Hz), 6.31 (1H, d, J=15.9 Hz).

Analysis: C₁₅H₁₂N₂O₅SCl₂×0.2H₂O, ×0.2 TFA Found: C=43.14%, H=3.04%,N=6.54%, S=7.19%. Calc.: C=43.05%, H=2.96%, N=6.52%, S=7.46%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.70 (1H, s), 10.65 (1H, s), 9.01 (1H, sbr), 7.91 (2H, d, J=8.4 Hz), 7.56 (2H, d, J=8.4 Hz), 7.45 (2H, d, J=8.1Hz), 7.33 (1H, d, J=15.6 Hz), 7.13 (2H, d, J=8.1 Hz), 6.31 (1H, J=15.6Hz).

Analysis: C₁₆H₁₃N₂O₅SF₃×0.2 TFA Found: C=46.43%, H=3.33%, N=6.22%,S=7.25%. Calc.: C=46.33%, H=3.13%, N=6.59%, S=7.54%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.66 (1H, s br), 10.37 (1H, s br), 8.56(1H, s br), 7.69 (2H, d, J=8.7 Hz), 7.39 (2H, d, J=8.1 Hz), 7.30 (1H, d,J=16.2 Hz), 7.10-7.03 (4H, m), 6.27 (1H, d, J=15.9 Hz), 3.77 (3H, s).

Analysis: C₁₆H₁₆N₂O₅S×0.7H₂O Found: C=53.32%, H=5.05%, N=7.98%, S=7.78%.Calc.: C=53.24%, H=4.86%, N=7.76%, S=8.88%.

¹H NMR: (300 MHz, DMSO d₆): δ=10.70 (1H, s), 10.66 (1H, s), 8.99 (1H,s), 8.06-7.98 (3H, m), 7.84-7.79 (1H, m), 7.45 (2H, d, J=8.4 Hz), 7.33(1H, d, J=15.6 Hz), 7.12 (2H, d, J=8.7 Hz), 6.32 (1H, d, J=15.9 Hz).

Analysis: C₁₆H₁₃F₃N₂O₄S Found: C=49.64%, H=3.30%, N=7.18%. Calc.:C=49.74%, H=3.39%, N=7.25%

¹H NMR: (300 MHz, DMSO d₆): δ=10.69 (1H, s, br), 10.47 (1H, s, br), 8.98(1H, s, br), 7.62 (1H, s), 7.58-7.56 (1H, m), 7.44-7.41 (4H, m), 7.32(1H, d, J=16.2 Hz), 7.11 (2H, d, J=8.1 Hz), 6.30 (1H, d, J=15.6 Hz),2.34 (3H, s).

Analysis: C₁₆H₁₆N₂O₄S×0.3 TFA Found: C=54.64%, H=4.75%, N=7.92%. Calc.:C=54.66%, H=4.59%, N=7.82%

¹H NMR: (300 MHz, MeOD d₄): 7.62-6.61 (m, 13H); 3.81 (broad s, 3H, OCH₃), 3.80 (broad s, 3H, OCH ₃),3.26 (broad s, 4H, NH).

¹³C NMR: (75 MHz, MeOD d₄): 167.0 (C═O); 154.4; 150.5; 143.1; 141.9;141.0; 132.5; 132.3; 129.9; 128.2; 126.7; 125.2; 122.4; 121.8; 120.8;119.6; 118.7; 111.9; 110.9; 56.6 (2C, OCH₃).

Combustion analysis: Calc: 60.91% C, 5.11% H, 9.27% N, 7.07% S.

Found: 60.40% C, 5.21% H, 9.16% N, 6.47% S.

HRMS: Calc: 453.1358. Found: 453.1351.

¹H NMR: (Acetone-d₆): δ (ppm): 9.25 (bs, 1H), 8.77 (bs, 1H), 7.79 (d,J=8.5 Hz, 2H), 7.61-7.51 (m, 5H), 7.36-7.28 (m, 3H), 6.99-6.93 (m, 1H),6.86-6.82 (m, 2H), 6.68-6.62 (m, 1H), 4.63 (bs, 2H).

HRMS: 449.1773 (calc.): 449.1767±0.0013 (found).

¹H NMR: (300 MHz, MeOD d₄): 8.00-6.56 (m, 13H); 3.77 (broad s, 3H, OCH₃), 3.74 (broad s, 3H, OCH ₃), 3.33 (broad s, 2H, NH), 3.00 (broad s,1H, NH), 2.88 (broad s, 1H, NH).

¹³C NMR: (75 MHz, MeOD d₄): 166.2 (C═O); 150.7; 148.5; 143.2; 141.7;140.6; 140.5; 131.9; 129.2; 128.9; 128.4; 126.7; 124.9; 119.5; 118.6;116.4; 113.2; 108.9; 56.6 (OCH₃); 56.4 (OCH₃).

MS: Calc: 453.1358. Found: 453.1351.

¹H NMR: (CD₃OD) δ (ppm): 7.68 (d, J=8.2 Hz, 2H), 7.55 (d, J=15.9 Hz,1H), 7.47 (d, J=8.5 Hz, 2H), 7.30 (d, J=8.0 Hz, 2H), 7.19-7.12 (m, 3H),7.03 (t, J=7.1 Hz, 1H), 6.86 (d, J=8.0 Hz, 1H), 6.75-6.69 (m, 2H), 2.37(s, 3H).

HRMS: 407.1304 (calc.): 407.1293±0.0012 (found).

¹H NMR: (300 MHz, DMSO-d₆) δ 10.6 (s, OH); 9 (s, NH); 7.1-7.8 (m, 14H,CH Ar); 6.2 (d, 1H, J=15 Hz)

¹H NMR: (300 MHz, MeODd₄): 7.31-6.62 (m, 11H); 3.72 (broad s, 3H); 3.70(broad s, 3H); 2.91 (t, 2H; J=7.1 Hz); 2.65 (broad t, 2H, J=7.4 Hz)

¹³C NMR: (75 MHz, MeODd₄): 173.9; 154.0; 150.3; 143.4; 138.6; 137.4;132.6; 130.2; 128.4; 127.4; 124.6; 123.1; 122.3; 119.3; 118.1; 111.7;110.9; 56.5 (2C); 38.8; 32.2.

HRMS: calc: 455.1515. Found: 455.1521.

¹H NMR: (300 MHz, DMSO d₆): 7.77 (d, 2H, J=8.8 Hz); 7.51 (d, 2H, J=8.5Hz); 7.34 (d, 2H, J=8.8 Hz); 7.18 (d, 2H, J=8.5 Hz); 7.11 (d, 2H, 8.8Hz); 6.94 (t, 1H, J=7.4 Hz); 6.77 (broad d, 2H, J=7.9 Hz); 6.6 (t, 1H,J=7.4 Hz), 4.95 (broad s, 1H), 3.83 (s, 3H).

¹³C NMR: (75 MHz, DMSO d₆): 162.5; 141.5; 139.2; 138.8; 130.9; 130.2;128.9; 128.6; 125.7; 124.7; 119.4; 116.2; 115.9; 114.5; 55.6.

HRMS: Calc: 423.1253. Found: 423.1235.

¹H NMR: (300 MHz, DMSO-d₆) δ 7.1-7.8 (m, 14H, CH Ar); 6.8-6.9 (m, 4H, CHAr); 6.3 (d, 1H, J=15 Hz)

Example 32

Sulfonamide 124 was prepared by condensation of 4-iodoaniline withbenzenesulfonyl chloride. Compound 125 was quantitatively furnished by aPd—Cu catalyzed coupling reaction of 124 with propargyl alcohol in basicsolvent. Primary alcohol 125 was oxidized to the correspondingcarboxylic acid 127 in two steps, including Dess-Martin periodinaneoxidation to afford aldehyde 126, followed by treatment with sodiumchlorite in buffered aqueous media in the presence of a chlorinescavenger. Acid 127 was derivatized to the hydroxamic acid 128 bytreatment with hydroxylamine hydrochloride and the coupling reagent EDCin the presence of N-hydroxybenzotriazole in basic, aprotic media.Compound 129 was prepared by coupling acid 130 with o-phenylenediamineas described in Example 31 for compound 86.

Data for 128:

¹H NMR: (300.07 MHz; acetone-d₆) δ (ppm): 9.4 (bs, 2H); 7.93 (dd, J=1.9,6.6; 2H); 7.82 (dd, J=1.9, 6.6; 2H); 7.68 (dd, J=1.4, 8.2; 2H); 7.48-741(m, 5H); 7.35-7.32 (m, 2H); 2.90 (bs, 1H)

¹³C NMR: (75.5 MHz; acetone-d₆) δ (ppm): 153.5; 147.2; 141.3; 140.3;139.5; 134.6; 130.1; 129.5; 128.8; 128.6; 128.3; 120.8; 116.5; 87.7;81.0.

MS: calc for C₂₁H₁₆O₄N₂S: 392.438. found: 393.4 for [M+H] (lowresolution MS).

Data for 129:

¹H NMR: (300.07 MHz; acetone-d₆) δ (ppm): 9.43 (bs, 1H); 8.02 (d, J=8.5Hz; 2H); 7.93 (d, J=8.5 Hz; 2H); 7.90 (d, J=8.5 Hz; 2H); 7.65 (d, J=8.5Hz; 2H); 7.47-7.34 (m, 7H); 7.21-7.17 (m, 2H); 2.80 (bs, 3H)

¹³C NMR: (75.5 MHz; acetone-d₆) δ (ppm): 167.2; 158.6; 146.3; 141.3;140.9; 139.8; 139.5; 134.2; 131.0; 129.9; 129.8; 129.3; 128.7; 128.6;128.4; 128.0; 126.8; 125.1; 122.7; 122.6; 120.1

MS: calc for C₂₇H₂₁O₃N₃S: 467.552. found: 468.5 for [M+H] (lowresolution MS).

Example 33

Benzylic alcohol 130 was prepared in 53% yield by addition of2-lithiofuran to styrene oxide. After protection of the resultinghydroxyl group with tert-butyldimethylsilyl chloride, the lithiatedspecies of compound 131 was treated with DMF to afford the formylderivative 132. Wadsworth-Horner-Emmons olefination was effected bytreatment of 132 with the sodium enolate of trimethylphosphonoacetate toafford the key intermediate 133 in 90% overall yield for the last threesteps. Saponification of the methyl ester with LiOH yielded the acid134, which in turn was converted into its hydroxamic acid form 135 byconventional activation with HOBt/EDC, followed by reaction withhydroxylamine Fluoride-promoted cleavage of silylated ether gave alcohol136 in 67% yield.

Data for 136:

¹H NMR: (300.07 MHz; acetone-d6) δ (ppm): 9.35 (bs, 1H); 7.40-7.15 (m;6H); 6.56 (d, J=2.9 Hz, 1H); 6.24 (d, J=15.3 Hz, 1H); 4.96 (t, J=6.2 Hz,1H); 3.00 (d, J=6.2 Hz, 2H)

¹³C NMR: (75.5 MHz; CD₃OD) δ (ppm): 166.6; 156.6; 151.3; 145.2; 129.3;128.5; 126.9; 116.2; 114.5; 111.0; 73.6; 39.1

Example 34

Unsaturated ketoacid 138 was obtained from ester 133 in 73% overallyield after three consecutive steps, including saponification(LiOH/H₂O/MeOH/THF), desilylation (TBAF/THF), and oxidation of benzylicalcohol 137 using Dess-Martin periodinane. Anilide 139 was obtained byBOP-mediated condensation of compound 138 with o-phenylenediamine in 83%yield.

Regioselective hydrogenation of the acrylate moiety in 133 wasaccomplished by treatment with NaBH₄ in the presence of NiCl₂, to affordthe propionate 140 in high yield. Ketoacid 142 was then obtained in 31%overall yield from 140 by an identical procedure to that followed in thesynthesis of 138 from 133. With compound 142 in hand, anilide 144 wasobtained as described above (BOP/o-phenylendiamine). The low yield wasdue to a difficult purification process. To avoid oxime formation,hydroxamic acid 143 was synthesized from 142 in 73% overall yield overtwo steps, including BOP-mediated coupling withN,O-bistrimethylsilylhydroxylamine, followed by cleavage of silylatedgroups under acidic conditions (citric acid/MeOH).

Data for 139:

¹H NMR: (300.07 MHz; CDCl₃) δ (ppm): 8.02-7.42 (series of multiplets,7H); 7.34 (bs, 1H); 7.06 (m, 1H); 6.80 (d, J=7.8; 1H); 6.79 (d, J=8.1;1H); 6.54 (d, J=3.0 Hz, 1H); 6.38 (m, 1H); 6.34 (d, J=3.0 Hz, 1H); 4.37(s, 2H); 3.90 (bs, 2H)

¹³C NMR: (75.5 MHz; CDCl₃) δ (ppm): 194.5; 164.4; 150.9; 150.8; 150.5;140.5; 135.9; 133.7; 128.7; 128.5; 126.9; 125.0; 124.4; 119.4; 118.0;117.5; 115.7; 111.3; 38.5

Data for 143:

¹H NMR: (300.07 MHz; CDCl₃) δ (ppm): 8.99 (bs, 1H); 8.09-7.42 (series ofmultiplets, 5H); 6.09 (d, J=3.0 Hz, 1H); 6.00 (d, J=3.0 Hz, 1H); 4.35(s, 2H); 2.95 (t, J=6.60 Hz, 2H); 2.50 (t, J=3.0 Hz, 1H).

¹³C NMR: (75.5 MHz; CDCl₃) δ (ppm): 196.2; 162.8; 153.2; 146.8; 134.9;133.7; 128.7; 128.5; 109.3; 107.1; 38.2; 31.7; 24.2

Data for 144:

¹H NMR: (300.07 MHz; CDCl₃) δ (ppm): 7.99-7.42 (series of multiplets,5H); 7.36 (bs, 1H); 7.02 (d, J=7.8, 2H); 6.73 (d, J=7.8 Hz, 2H); 6.13(d, J=3.0 Hz, 1H); 6.04 (d, J=3.0 Hz, 1H); 4.30 (s, 2H); 3.70 (bs, 2H);3.03 (t, J=6.9 Hz, 2H); 2.69 (t, J=6.9 Hz, 2H).

¹³C NMR: (75.5 MHz; CDCl₃) δ (ppm): 195.4; 170.7; 153.6; 147.1; 140.9;136.1; 133.5; 128.7; 128.5; 127.1; 125.7; 124.0; 119.2; 117.8; 109.1;107.2; 38.4; 35.7; 24.7

Example 35 General Procedure for Synthesis of Urea Compounds

To a solution of isocyanate (1.5 mmol) in 15 mL of anhydrousdichloromethane, was added a solution of 4-anilinylmethylacrylate (1.5mmol) in dichloromethane (10 mL). The mixture was stirred at roomtemperature for 15 hours. After addition of ammonium chloride solutionthe new mixture was extracted from dichloromethane. The organic layerswere combined and washed with ammonium chloride solution, water, brineand dried over magnesium sulfate. The crude was then flashed over silicagel using CH₂Cl₂: MeOH (9.5:0.5) as eluent.

The following compounds were synthesized according to the generalprocedure:

¹H NMR: (300 MHz, DMSO-d₆) δ 7.5-7.7 (m, 4H, CH Ar); 7.5 (d, 2H, J=6.6Hz); 7.3 (d, 2H, J=6.6 Hz); 6.3 (d, 1H, J=15 Hz)

¹H NMR: (300 MHz, DMSO-d₆) δ 7.5-8.2 (m, 7H, CH Ar); 7.5 (d, 2H, J=6.6Hz); 7.3 (d, 2H, J=6.6 Hz); 6.3 (d, 1H, J=15 Hz)

¹H NMR: (300 MHz, DMSO-d₆) δ 7.5-7.7 (m, 3H, CH Ar); 7.5 (d, 2H, J=6.6Hz); 7.3 (d, 2H, J=6.6 Hz); 6.3 (d, 1H, J=15 Hz)

Example 36

The following additional compounds were prepared by procedures analogousto those described in the foregoing Examples:

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 1.99 (m, 2H), 2.79 (t, 2H, J=7.2 Hz),3.21 (dd, 2H, J=6.8, 7.8 Hz), 7.27 (d, 2H, J=8.1 Hz), 7.65 (t, 2H, J=7.8Hz), 7.72-7.77 (m, 3H), 7.90 (d, 2H, J=7.2 Hz), 10.77 (broad s, 1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 25.2 (t), 34.3 (t), 55.6 (t), 128.0(d), 2×128.8 (d), 129.4 (d), 2×130.2 (d), 131.1 (s), 134.5 (d), 140.7(s), 145.5 (s), 165.8 (s).

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 1.66-1.88 (m, 4H), 2.71 (t, 2H, J=6.3Hz), 4.34 (d, 1H, J=303 Hz), 4.87 (m, 1H), 7.27 (d, 2H, J=7.8 Hz),7.44-7.48 (m, 2H), 7.52 (dd, 1H, J=1.5, 9.4 Hz), 7.73 (d, 2H, J=7.8 Hz),7.83 (s, 1H), 7.83-7.88 (m, 3H), 8.16 (broad s, 1H), 10.67 (broad s,1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 28.3 (t), 36.2 (t), 39.8 (t), 74.0 (d),125.0 (d), 125.3 (d), 126.2 (d), 126.7 (d), 2×127.8 (d), 128.4 (d),128.5 (d), 128.6 (d), 2×129.3 (d), 130.6 (s), 133.7 (s), 134.3 (s),144.7 (s), 147.4 (s), 165.9 (s).

¹H NMR: (300 MHz, DMSO-d6) δ 11.2 (s, OH); 9 (s, NH); 7.6-7.8 (m, 4H, CHAr); 7-7.4 (m, 5H, CH Ar); 2.8 (m, 4H, CH₂).

¹H NMR: (300 MHz, DMSO-d₆) δ 11.2 (s, 1H); 9.0 (s, 1H); 7.7 (m, 6H);7.34 (m, 5H).

¹H NMR: (300 MHz, DMSO-d₆) δ 11.2 (s, OH); 9 (s, NH); 7.6-7.8 (m, 4H, CHAr); 7-6.8 (m, 4H, CH Ar); 2.9 (s, 6H, 2CH₃); 2.8 (m, 4H, CH₂).

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 1.38 (quintuplet, 2H, J=7.5 Hz),1.60-1.72 (m, 4H), 2.60 (t, 2H, J=7.8 Hz), 2.67 (t, 2H, J=7.5 Hz),7.15-7.31 (m, 7H), 7.75 (d, 2H, J=8.1 Hz), 8.11 (broad s, 1H), 10.68(broad s, 1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 31.8 (t), 32.1 (t), 36.2 (t), 36.4 (t),126.4 (d), 127.8 (d), 2×129.0 (d), 2×129.2 (d), 2×129.3 (d), 143.3 (s).

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 1.63 (m, 4H, J=4.5 Hz), 2.37 (t, 2H,J=7.8 Hz), 2.57-2.66 (m, 4H), 2.86 (t, 2H, J=7.5 Hz), 7.10-7.28 (m, 9H),8.01 (broad s, 1H), 9.98 (broad s, 1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 31.0 (t), 2×31.9 (t), 35.1 (t), 35.8(t), 36.2 (t), 126.4 (d), 2×129.0 (d), 2×129.1 (d), 2×129.1 (d), 129.2(d), 138.8 (s), 141.2 (s), 143.4 (s), 164.1 (s).

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 1.83-1.98 (m, 4H), 2.08-2.14 (m, 2H),2.56-2.67 (m, 6H), 7.12-7.30 (m, 9H), 9.98 (broad s, 1H).

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 1.60-1.68 (m, 4H), 1.87 (quintuplet,2H, J=7.5 Hz), 2.03-2.14 (m, 2H), 2.55-2.67 (m, 6H), 7.09-7.28 (m, 9H).

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 2.37 (t, 2H, J=7.2 Hz), 2.78-2.89 (m,6H), 7.13-7.29 (m, 9H), 7.84 (broad s, 1H), 9.90 (broad s, 1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 31.6 (t), 35.1 (t), 38.2 (t), 38.6 (t),2×126.6 (d), 2×129.1 (d), 2×129.2 (d), 2×129.3 (d), 139.4 (s), 140.4(s), 142.8 (s), 170.1 (s).

¹H NMR (300.072 MHz, (CD₃)₂CO): δ 1.96 (quintuplet, 2H, J=6.0 Hz), 2.69(t, 2H, J=8.0 Hz), 3.19 (dd, 2H, J=6.0, 9.0 Hz), 3.38 (s, 2H), 7.09 (d,2H, J=7.5 Hz), 7.21 (d, 2H, J=7.5 Hz), 7.66 (t, 2H, J=8.1 Hz), 7.747 (t,1H, J=6.9 Hz), 7.90 (d, 2H, J=6.6 Hz), 10.08 (broad s, 1H).

¹³C NMR (75.46 MHz, (CD₃)₂CO): δ 25.5 (t), 34.1 (t), 39.9 (t), 55.7 (t),2×128.8 (d), 130.0 (d), 2×130.2 (d), 134.4 (s), 139.9 (s), 140.7 (s),168.5 (s).

¹H NMR: (300 MHz, DMSO d₆): δ 7.77 (broad s, 4H); 7.57 (d, 1H, J=15.7Hz); 7.35 (d, 1H, J=6.9 Hz); 7.03-6.94 (m, 6H); 6.76 (d, 1H, J=7.1 Hz);6.59 (d, 1H, J=6.9 Hz); 4.98 (broad s, 2H); 2.19 (s, 3H).

¹³C NMR: (75 MHz, DMSO d₆): δ 162.9; 141.6; 139.8; 139.0; 137.6; 134.8;133.6; 129.6; 128.1; 127.3; 125.9; 125.4; 124.7; 123.2; 120.7; 116.2;115.9; 20.3.

¹H NMR: (300 MHz, DMSO d₆): δ 7.91-7.81 (m, 4H); 7.63-7.58 (m, 5H);7.48-7.43 (m, 2H); 7.39-7.33 (m, 2H); 7.24 (d, 2H, J=8.5 Hz); 6.97 (dd,2H, J=9.9, 7.1 Hz); 6.79 (d, 1H, J=7.7 Hz) 6.61 (dd, 1H, J=7.7, 7.1 Hz);5.01 (broad s, 2H).

¹³C NMR: (75 MHz, DMSO d₆): δ 162.9; 141.9; 141.6; 139.8; 139.2; 137.6;136.9; 135.8; 128.9; 128.3; 127.4; 127.3; 127.2; 126.3; 126.0; 125.5;124.8; 123.2; 120.4; 116.2; 115.9.

¹H NMR: (300 MHz, MeODd₄): δ 7.74-7.54 (m, 5H); 7.07-6.96 (m, 4H); 6.55(d, 1H, J=15.7); 2.25 (s, 3H).

¹³C NMR: (75 MHz, MeODd₄): δ 163.5; 141.6; 140.4; 139.5; 136.1; 135.9;130.6; 129.0; 128.8; 123.1; 121.7; 20.8

¹H NMR: (300 MHz, MeODd₄): δ 7.83-7.19 (m, 14H); 6.56 (d, 1H, J=15.7Hz).

¹³C NMR: (75 MHz, MeODd₄): δ 165.4; 141.6; 141.4; 140.5; 139.5; 139.0;137.9; 129.8; 129.2; 128.7; 128.6; 128.2; 127.6; 122.7; 121.7.

Example 37

To a solution of carboxylic acid 163 (131 mg, 0.36 mmol), preparedaccording to procedures described above, in 6 mL of dry DMF was addedEt₃N (190 μl, 1.37 mmol), followed by the addition of solid BOP (259 mg;0.59 mmol). The reaction mixture was stirred for 10 min. at roomtemperature and then solid 5-amino-1,3,4-thiadiazole-2-thiol (58 mg,0.43 mmol) was added. After being stirred for 12 h, the mixture wasdiluted with methanol and concentrated under vacuum. Upon dilution withCH₂Cl₂/MeOH, crystallization of 164 (150 mg, 87%) from the crude oiltook place.

¹H NMR: (300 MHz, DMSO d₆): δ 7.85 (broad s, 5H); 7.04-6.58 (m, 4H);3.69 (s, 3H); 3.67 (s, 3H); 3.38 (broad s, 3H).

¹³C NMR: (75 MHz, DMSO d₆): 163.3; 161.7; 158.7; 148.7; 146.2; 142.0;140.7; 137.9; 130.1; 128.7; 127.5; 121.4; 113.7; 112.0; 106.6; 55.5;55.4.

Following this general procedure, the following thiadiazole derivativeswere prepared from the corresponding carboxylic acids:

¹H NMR: (300 MHz, DMSO-d₆); δ (ppm): 7.89-7.72 (series of multiplets,7H); 7.50-7.05 (series of multiplets, 6H); 3.32 (broad singlet, 3H)

¹³C NMR: (75 MHz, DMSO-d6); d (ppm): 162.6; 162.3; 144.5; 138.3; 138.3;138.2; 132.5; 130.1; 129.7; 129.1; 128.6; 127.6; 127.3; 127.1; 120.9;118.7; 116.8.

MS: calc. for M+H, 493.6. obs. for M+H, 496.3

¹H NMR: (300 MHz, DMSO d₆): 7.87-7.72 (m, 5H), 7.57-7.53 (m, 4H), 7.39(dd, 2H, J=6.9, 7.7 Hz), 7.30 (d, 1H, J=7.1 Hz), 7.17 (d, 2H, J=8.5 Hz),6.85 (d, 1H, J=15.9 Hz).

MS: cal: 495.61. found: 496.6.

Following an analogous procedure, but substituting2-amino-5-trifluoro-methyl-1,3,4-thiadiazole for5-amino-1,3,4,-thiadiazol-2-thiol, the following compound was prepared:

¹H NMR: (300 MHz, DMSO d₆): δ 7.96-7.81 (m, 5H); 7.71-7.48 (m, 4H); 7.38(dd, 2H, J=7.1, 7.41 Hz); 7.28 (d, 1H, J=7.1 Hz); 7.19 (d, 2H, J=8.5Hz); 6.98 (d, 1H, J=15.7 Hz).

¹³C NMR: (75 MHz, DMSO d₆): 192.3; 163.6; 161.6; 142.4; 140.9; 139.2;138.0; 136.8; 135.9; 129.0; 128.8; 127.4; 127.2; 126.2; 121.2; 120.4.

MS: cal: 530.55 found: 531.5.

Example 38

Coupling of 24 (from Example 15) with o-phenylenediamine in the presenceof benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (BOP) afforded the anilide 168.

By an analogous procedure, the corresponding para-substituted compoundis prepared from 32 (from Example 16).

Example 39

Step 1: N-Methyl-4-iodophenylbenzenesulfonamide (169)

To compound 28 (from Example 18) (500 mg, 1.39 mmol) in DMF (10 mL) wereadded at room temperature K₂CO₃ (962 mg, 6.96 mmol), followed by methyliodide (395 mg, 2.78 mmol). The resulting reaction mixture was stirredat room temperature for 16 hours. The solvent is then removed and waterwas added. The resulting mixture was extracted with ethyl acetate, andthe combined organic phases were dried and concentrated. Purification byflash chromatography using hexane:ethyl acetate (8:2) afforded 510 mg(98%) of the title compound as a white solid.

Compound 169 was converted to the hydroxamic acid 170 according to theprocedures described in Example 18 for the preparation of compound 36.

Data for 170:

¹H NMR: (300 MHz, DMSO d₆): δ=10.76 (1H, s), 9.04 (1H, s), 7.73-7.68(1H, m), 7.61-7.51 (6H, m), 7.43 (1H, d, J=15.9 Hz), 7.15 (2H, d, J=8.7Hz), 6.43 (1H, d, J=16.2 Hz), 3.15 (3H, s).

Analysis: C₁₆H₁₆N₂O₄S×0.5H₂O Found: C=56.36%, H=5.09%, N=8.69%, S=8.33%.Calc.: C=56.29%, H=5.02%, N=8.21%, S=9.39%.

Example 40 N-hydroxy-2-(4-(4-phenylbutyl)phenyl)acetamide (174) Step 1:Methyl 2-(4-iodophenyl)acetate

A 4M solution of HCl in dioxane (50 mL) was added to2-(4-iodophenyl)acetic acid (10 g, 38.2 mmol) in MeOH (100 mL) and thereaction stirred overnight. The solvent was evaporated under reducedpressure. The residue was purified by silica gel column chromatographyeluting with 0-30% EtOAc in hexanes to afford methyl2-(4-iodophenyl)acetate in 10.42 g (99%). LRMS: 276.0 (calc) 277.1(found) (MH)⁺.

Step 2: Methyl 2-(4-(4-phenylbut-1-ynyl)phenyl)acetate

CuI (0.06 equiv, 207 mg, 1.1 mmol) was added to a solution of Pd(Ph₃P)₄(0.03 equiv, 628 mg, 0.54 mmol) and aromatic iodide (1 equiv, 5 g, 18mmol) in Et₂NH:DME (30 mL:30 mL) and the reaction stirred for 20 min.4-phenyl-1-butyne (3 equiv, 7.1 g, 54 mmol) was then added dropwise andthe reaction stirred for 3 h. The reaction was concentrated underreduced pressure and then the residue partitioned between EtOAc (50 mL)and H₂O (50 mL). The organic phase was separated, dried over Na₂SO₄filtered and concentrated. The compound was purified by silica gel flashcolumn chromatography: 20%-100% EtOAc in hexanes to afford methyl2-(4-(4-phenylbut-1-ynyl)phenyl)acetate in 4.95 g (98%). LRMS: 278.3(calc) 279.2 (found) (MH)⁺.

Step 3: Methyl 2-(4-(4-phenylbutyl)phenyl)acetate

10% Pd/C (20% w/w, 1 g) was added to a solution of the acetylene (4.9 g,18 mmol) in MeOH (30 mL). The reaction was then purged with H₂ andstirred overnight. The solvent was evaporated and the residue waspurified by a silica plug eluting with 30% EtOAc in hexanes to affordmethyl 2-(4-(4-phenylbutyl)phenyl)acetate in 4.99 g (99%). LRMS: 282.3(calc) 283.1 (found) (MH)⁺.

Step 4: N-hydroxy-2-(4-(4-phenylbutyl)phenyl)acetamide (174)

NaOH (6 equiv, 4.2 g, 106 mmol) was added to a solution of the methylester (1 equiv, 4.9 g, 18 mmol) and aq. NH₂OH (50 equiv, 58 g, 884 mmol)in MeOH (100 mL) and THF (100 mL) at 23° C. After stirring the reactionovernight, the reaction was adjusted to pH=7. The solvent was evaporatedand the residue was purified by trituration with hexanes and water toafford N-hydroxy-2-(4-(4-phenylbutyl)phenyl)acetamide 174 in 4.66 g(93%). (dDMSO) δ (ppm) ¹H, 10.59 (s, 1H), 8.77 (s, 1H), 7.25-7.06 (m,9H), 3.19 (s, 2H), 2.58-2.53 (m, 4H), 1.54-1.52 (m, 4H). LRMS: 283.1(calc) 282.0 (MH)⁻.

Example 41N-hydroxy-2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetamide (179)Step 1: Methyl 2-(4-(3-hydroxyprop-1-ynyl)phenyl)acetate

Following the procedure of Step 2 of Example 40 above afforded methyl2-(4-(3-hydroxyprop-1-ynyl)phenyl)acetate in 1.17 g (79%) as a thickyellow oil. (MeOD-d4) δ (ppm) ¹H, 7.36 (d, J=8.4 Hz, 2H), 7.24 (d, J=8.0Hz, 2H), 4.38 (s, 2H), 3.67 (s, 3H), 3.65 (s, 2H).

Step 2: Methyl 2-(4-(3-hydroxypropyl)phenyl)acetate

Following the procedure of Step 3 of Example 40 above afforded methyl2-(4-(3-hydroxypropyl)phenyl)acetate in 1.18 g (99%) a clear translucentoil. (MeOD-d4) δ (ppm) ¹H, 7.15 (d, J=1.6 Hz, 4H), 3.66 (s, 3H), 3.59(s, 2H), 3.55 (t, J=6.4 Hz, 2H), 2.65 (t, J=7.6 Hz, 2H), 1.82 (m, 2H).

Step 3: Methyl 2-(4-(3-oxopropyl)phenyl)acetate

TEMPO (0.02 equiv, 6 mg, 0.038 mmol) was added to a solution of methyl2-(4-(3-hydroxypropyl)phenyl)acetate in CH₂Cl₂ (5 mL) and cooled to 0°C. KBr (2.2 equiv, 1.6 mL, 2.7M) and KHCO₃ (5.5 equiv, 6.6 mL, 1.6M)solutions were then added followed by the dropwise addition of 10% aq.NaOCl (1.34 equiv, 1.9 g, 2.6 mmol). Once the addition was complete, thereaction was stirred for an additional 10 min. at 0° C. The reaction wasquenched with sat. aq.sodium thiosulfate solution (3 mL) and thenpartitioned between H₂O (10 mL) and CH₂Cl₂ (10 mL). The organic phasewas separated and the remaining aqueous phase was extracted with EtOAc(2×2 mL). The organic layers were combined, dried over Na₂SO₄, filteredand concentrated under reduced pressure. The resulting aldehyde wascarried forward to the subsequent reaction without further purification.LRMS: 206.1 (calc) 207.2 (found) (MH)⁺.

Step 4: Methyl 2-(4-(but-3-ynyl)phenyl)acetate

Dimethyl-2-oxopropylphosphonate (1.2 equiv, 1.5 g, 9.1 mmol) was addedto a suspension of K₂CO₃ (3 equiv, 3.1 g, 23 mmol) and p-TsN₃ (1.2equiv, 1.8 g, 9.1 mmol) in MeCN (144 mL). The mixture was then stirredfor 2 h. A solution of the methyl 2-(4-(3-oxopropyl)phenyl)acetate (1equiv, 1.56 g, 7 6 mmol) in MeOH was then added in one portion and thereaction was stirred overnight. The solvent was evaporated under reducedpressure. The residue was partitioned between Et₂O (100 mL) and water(100 mL). The aqueous phase was separated and extracted with Et₂O (25mL) twice. The organic phases were combined, dried over Na₂SO₄, filteredand concentrated. The residue was purified by silica gel flash columneluting with 0-30% EtOAc in hexanes to afford methyl2-(4-(but-3-ynyl)phenyl)acetate in 980 mg (64%). LRMS: 202.5 (calc)225.1 (found) (MNa)⁺.

Step 5: Methyl 2-(4-(4-(2,4,5-trifluorophenyl)but-3-ynyl)phenyl)acetate

Following the procedure of Step 2 of Example 40 above afforded methyl2-(4-(4-(2,4,5-trifluorophenyl)but-3-ynyl)phenyl)acetate in 95 mg (39%)as a yellow oil. LRMS: 332.3 (calc) 355.3 (found) (MNa)⁺.

Step 6: Methyl 2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetate

Following the procedure of Step 3 of Example 40 above afforded methyl2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetate in 91 mg (95%) as aclear translucent oil. LRMS: 336.3 (calc) 359.2 (found) (MNa)⁺.

Step 7: N-hydroxy-2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetamide(179)

Following the procedure of Step 4 of Example 40 above affordedN-hydroxy-2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetamide 179 in52 mg (57%) as a white powder. (MeOD-d4) δ (ppm) ¹H, 7.15 (m, 6H), 3.35(s, 2H), 2.61 (m, 4H), 1.60 (m, 4H). LRMS: 337.3 (calc) 338.3 (found)(MH)⁺.

Example 422-(4-(4-(benzo[c][1,2,5]oxadiazol-5-yl)but-3-ynyl)phenyl)-N-hydroxyacetamide(180)

Following the procedure of Step 4 of Example 40 above afforded2-(4-(4-(benzo[c][1,2,5]oxadiazol-5-yl)but-3-ynyl)phenyl)-N-hydroxyacetamide180 in 8 mg (57%) as a yellowish orange powder. (MeOD-d4) δ (ppm) ¹H,10.62 (s, 1H), 8.79 (s, 1H), 8.07 (s, 1H), 8.03 (d, J=9.2 Hz, 1H), 7.40(d, J=9.6 Hz, 1H), 7.24 (d, J=8.4 Hz, 2H), 7.18 (d, J=8.0 hz, 2H), 3.23(s, 3H), 2.83 (m, 2H), 2.77 (m, 2H) LRMS: 321.1 (calc) 320.3 (found)(MH)⁻.

Example 43 N-Hydroxy-2-(3-(4-phenylbutyl)phenyl)acetamide (181) Step 1:Methyl 2-(3-(4-hydroxybut-1-ynyl)phenyl)acetate

Following the procedure of Step 2 of Example 40 above afforded methyl2-(3-(4-hydroxybut-1-ynyl)phenyl)acetate in 227 mg (34%) as a yellowoil. LRMS: 218.2 (calc) 219.1 (found) (MH)⁺.

Step 2: Methyl 2-(3-(4-hydroxybutyl)phenyl)acetate

Following the procedure of Step 3 of Example 40 above afforded methyl2-(3-(4-hydroxybutyl)phenyl)acetate in 181 mg (78%) as a cleartranslucent oil. LRMS: 222.3 (calc) 223.1 (found) (MH)⁺.

Step 3: Methyl 2-(3-(4-oxobutyl)phenyl)acetate

Following the procedure of Step 3 of Example 41 above afforded methyl2-(3-(4-oxobutyl)phenyl)acetate in 178 mg (99%) as a clear translucentoil. LRMS: 220.3 (calc) 221.4 (found) (MH)⁺.

Step 4: Methyl 2-(3-(4-hydroxy-4-phenylbutyl)phenyl)acetate

A 1.0 M solution of PhMgBr (1 equiv, 0.45 mmol) was added dropwise to asolution of methyl 2-(3-(4-oxobutyl)phenyl)acetate (1 equiv, 100 mg,0.45 mmol) in THF (2 mL) at 0° C. The reaction was then allowed to warmto 23° C. over 1 h. The reaction was quenched by the addition on sat.aq. NH₄Cl (10 mL). The aqueous phase was separated and extracted withEtOAc (2×5 mL). The organic layers were then combined, dried overNa₂SO₄, filtered and concentrated under reduced pressure. The crudeproduct was purified by silica gel column chromatography eluting with0-50% EtOAc in hexanes to afford methyl2-(3-(4-hydroxy-4-phenylbutyl)phenyl)acetate in 78 mg(58%) as a cleartranslucent oil. LRMS: 298.4 (calc) 321.2 (found) (MH)⁺.

Step 5: Methyl 2-(3-(4-phenylbutyl)phenyl)acetate

Following the procedure of Step 3 of Example 40 above afforded methyl2-(3-(4-phenylbutyl)phenyl)acetate in 15 mg (20%) as a clear translucentoil. LRMS: 282.4 (calc) 283.2 (found) (MH)⁺.

Step 6: N-Hydroxy-2-(3-(4-phenylbutyl)phenyl)acetamide (181)

Following the procedure of Step 4 of Example 40 above affordedN-Hydroxy-2-(3-(4-phenylbutyl)phenyl)acetamide (181) in 5 mg (33%) as awhite powder. (CD₃OD) δ (ppm) 1H, 7.23 (m, 9H), 3.36 (s, 2H), 2.61 (m,4H), 1.63 (m, 4H) LRMS (ESI): (calc.) 283.1 (found) 282.2 (MH)⁻

Example 44

The following additional compounds were prepared by procedures analogousto those described in the Examples 40-43:

-   a)    N-hydroxy-2-(4-(4-(4-(trifluoromethyl)phenyl)butyl)phenyl)acetamide    (182)

(CD3OD) δ (ppm) 1H, 7.53 (d, J=8.4 Hz, 2H), 7.33 (d, J=8.0 Hz, 2H), 7.18(d, J=8.0 Hz, 2H), 7.10 (d, J=8.0 Hz, 2H), (s, 2H), 2.70 (t, J=7.2 Hz,2H), 2.61 (t, J=7.2 Hz, 2H), 1.63 (m, 4H). LRMS (ESI): (calc.) 351.14(found) 350.24 (MH)⁻.

-   b) 2-(4-(4-(1H-indol-5-yl)butyl)phenyl)-N-hydroxyacetamide (183)

(CD3OD) d(ppm) 1H, 7.29 (m, 1H), 7.24 (d, J=8.8 Hz, 1H), 7.18-7.16 (m,3H), 7.11-7.09 (m, 2H), 6.90 (dd, J=8.4 Hz, 1.6 Hz, 1H), 6.34 (d, J=4Hz, 1H), 3.35 (s, 2H), 2.68 (t, J=7.2 Hz, 2H), 2.61 (t, J=7.2 Hz, 2H),1.64 (m, 4H) LRMS (ESI): (calc.) 322.17 (found) 323.421 (MH)⁺.

-   c)    N-hydroxy-2-(4-(4-(3-(trifluoromethyl)phenyl)butyl)phenyl)acetamide    (184)

(dDMSO) δ (ppm) 1H, 10.60 (s, 1H), 8.77 (s, 1H), 7.50 (m, 4H), 7.12 (d,J=8.0 Hz, 2H), 7.07 (d, J=8.0 Hz, 2H), 3.19 (s, 2H), 2.68 (t, J=7.2 Hz,2H), 2.54 (t, J=7.2 Hz, 2H), 1.55 (m, 4H). LRMS (ESI): (calc.) 351.1(found) 350.3 (MH)⁻.

-   d)    N-hydroxy-2-(4-(4-(2-(trifluoromethyl)phenyl)butyl)phenyl)acetamide    (185)

(dDMSO) d(ppm) 1H, 10.60 (s, 1H), 8.77 (s, 1H), 7.63 (d, J=8.0 Hz, 1H),7.57 (t, J=7.2 Hz, 1H), 7.43 (d, J=7.6 Hz, 1H), 7.37 (t, J=8.0 Hz, 1H),7.13 (d, J=8.0 Hz, 2H), 7.09 d, J=8.0 Hz, 2H), 3.20 (s, 2H), 2.73 (t,J=7.6 Hz, 2H), 1.58 (m, 4H). LRMS (ESI): (calc.) 351.1 (found) 350.3(MH)−.

-   e)    N-hydroxy-2-(4-(4-(imidazo[1,2-a]pyridin-6-yl)butyl)phenyl)acetamide    (186)

(CD3OD) d(ppm) 1H, 8.19 (s, 1H), 7.73 (s, 1H), 7.49 (s, 1H), 7.43 (d,J=9.2 Hz, 1H), 7.15 (m, 5H), 3.34 (s, 2H), 2.63 (m, 4H), 1.66 (m, 4H).LRMS (ESI): (calc.) 323.1 (found) 324.3 (MH)+

-   f)    2-(4-(4-(benzo[d][1,3]dioxol-5-yl)butyl)phenyl)-N-hydroxyacetamide    (187)

(dDMSO) d(ppm) 1H, 10.60 (s, 1H), 8.78 (s, 1H), 7.12 (d, J=8.0 Hz, 2H),7.07 (d, J=8.0 Hz, 2H), 6.75 (m, 2H), 6.60 (d, J=8.0 Hz, 1H), 5.92 (s,2H), 3.19 (s, 2H), 2.52 (m, 4H), 1.50 (m, 4H). LRMS (ESI): (calc.) 327.1(found) 326.4 (MH)−.

-   g)    2-(4-(4-(benzo[d][1,3]dioxol-5-yl)but-3-ynyl)phenyl)-N-hydroxyacetamide    (188)

(dDMSO) d(ppm) 1H, 10.61 (s, 1H), 8.78 (s, 1H), 7.20 (d, J=8.4 Hz, 2H),7.16 (d, J=8.4 Hz, 2H), 6.85 (m, 3H), 6.01 (s, 2H), 3.22 (s, 2H), 2.77(t, J=7.6 Hz, 2H), 2.61 (t, J=6.8 Hz, 2H). LRMS (ESI): (calc.) 323.1(found) 322.2 (MH)−.

-   h)    2-(4-(4-(benzo[c][1,2,5]oxadiazol-5-yl)but-3-ynyl)phenyl)-N-hydroxyacetamide    (189)

(dDMSO) d(ppm) 1H, 10.60 (s, 1H), 8.78 (s, 1H), 7.12 (d, J=8.0 Hz, 2H),7.07 (d, J=8.0 Hz, 2H), 6.75 (m, 2H), 6.60 (d, J=8.0 Hz, 1H), 5.92 (s,2H), 3.19 (s, 2H), 2.52 (m, 4H), 1.50 (m, 4H). LRMS (ESI): (calc.) 327.1(found) 326.4 (MH)−.

-   i) N-hydroxy-2-(4-(2-phenoxyethoxy)phenyl)acetamide (190)

(DMSO) d(ppm) 1H, 10.58 (bs, 1H), 8.77 (bs, 1H), 7.28 (t, J=7.6 Hz<2H),7.15 (d, J=8.8 Hz, 2H), 6.93 (m, 5H), 4.26 (s, 4H), 3.18 (s, 2H). LRMS(ESI): (calc.) 287.3 (found) 310.0 (MH)⁺.

-   j) N-hydroxy-2-(4-(3-phenoxypropyl)phenyl)acetamide (191)

(CD3OD) d(ppm) 1H, 7.21 (m, 6H), 6.88 (m, 6H), 3.92 (t, J=8 Hz, 2H),3.35 (s, 2H), 2.77 (t, J=7.2 Hz, 2H), 2.04 (m, 2H). LRMS (ESI): (calc.)285.3 (found) 286.2 (MH)⁺.

-   k) 2-(4-butylphenyl)-N-hydroxyacetamide (192)

(CD3OD) d(ppm) 1H, 7.18 (d, J=8 Hz, 2H), 7.11 (d, J=8 Hz, 2H), 3.35 (s,2H), 2.57 (T, J=7.6 Hz, 2H), 1.56 (m, 2H), 1.33 (m, 2H), 0.92 (t, J=7.2Hz, 2H). LRMS (ESI): (calc.) 207.2 (found) 208.1 (ES+;Na+).

-   l) N-hydroxy-2-(4-(5-phenylpentyl)phenyl)acetamide (193)

(CD3OD) d(ppm) 1H, 7.22-7.08 (m, 9H), 3.34 (s, 2H), 2.56 (m, 4H), 1.61(m, 4H), 1.34 (m, 2H). LRMS (ESI): (calc.) 297.4 (found) 298.3 (MH)⁺.

-   m) N-hydroxy-2-(4-(4-hydroxy-4-(pyridin-4-yl)butyl)phenyl)acetamide    (194)

(CD3OD) d(ppm) 1H, 8.44 (d, J=6.0 Hz, 2H), 7.36 (d, J=6.4 Hz, 2H), 7.18(d, J=8.4 Hz, 2H), 7.09 (d, J=8.0 Hz, 2H), 4.65 (m, 1H), 3.34 s, 2H),2.60 (m, 2H), 1.69 (m, 4H). LRMS (ESI): (calc.) 300.3 (found) 301.3(MH)⁺.

-   n) N-hydroxy-2-(4-(4-hydroxy-4-(pyridin-3-yl)butyl)phenyl)acetamide    (195)

(CD3OD) d(ppm) 1H, 8.46 (s, 1H), 8.41 (d, J=4.8 Hz, 1H), 7.78 (d, J=8.0Hz, 1H), 7.39 (t, J=5.6 Hz, 1H), 7.18 (d, J=8.0 Hz, 2H), 7.09 (d, J=8.0Hz, 2H), 4.69 (m, 1H), 3.34 (s, 2H), 2.61 ((t, J=6.4 Hz, 2H), 1.79-1.58(m, 4H). LRMS (ESI): (calc.) 300.5 (found) 301.4 (MH)+.

-   o) N-hydroxy-2-(4-(4-hydroxy-4-(pyridin-2-yl)butyl)phenyl)acetamide    (196)

(CD3OD) d(ppm) 1H, 8.42 (m, 1H), 7.81 (m, 1H), 7.51 (d, J=7.2 Hz, 1H),7.26 (d, J=4.8 Hz, 1H), 7.16 (m, 1H), 7.08 (m, 2H), 4.69 (m, 1H), 3.34(s, 2H), 2.59 (m, 2H), 1.76-1.69 (m, 4H). LRMS (ESI): (calc.) 300.3(found) 301.4 (MH)⁺.

-   p) N-hydroxy-2-(4-(4-(pyridin-4-yl)butyl)phenyl)acetamide (197)

(CD3OD) d(ppm) 1H, 8.37 (d, J=6.0 Hz, 2H), 7.24 (d, J=5.6 Hz, 2H), 7.18(d, J=8.0 Hz, 2H), 7.10 (d, J=8.0 Hz, 2H), 3.35 (s, 2H), 2.67 (t, J=6.4Hz, 2H), 2.62 (t, J=7.2 Hz, 2H), 1.65 (m, 4H). LRMS (ESI): (calc.) 284.1(found) 285.3 (MH)+.

-   q) N-hydroxy-2-(4-(4-(pyridin-3-yl)butyl)phenyl)acetamide (198)

(CD3OD) d(ppm) 1H, 8.24 (m, 2H), 7.65 (d, J=8.0 Hz, 1H), 7.33 (m, 1H),7.18 (d, J=7.6 Hz, 2H), 7.10 (d, J=7.6 Hz, 2H), 3.35 (s, 2H), 2.64 (m,4H), 1.64 (m, 4H). LRMS (ESI): (calc.) 284.1 (found) 285.3 (MH)+.

-   r) N-hydroxy-2-(4-(4-(pyridin-2-yl)butyl)phenyl)acetamide (199)

(CD3OD) d(ppm) 1H, 8.39 (d, J=6.0 Hz), 7.75 (t, J=8.0 Hz, 1H), 7.23 (m,4H), 7.10 (d, J=8.0 Hz, 2H), 3.34 (s, 2H), 2.78 (t, J=7.6 Hz, 2H), 2.61(t, J=7.6 Hz, 2H), 1.67 (m, 4H). LRMS (ESI): (calc.) 284.1 (found) 285.4(MH)+.

-   s) N-hydroxy-2-(4-(3-phenylpropyl)phenyl)acetamide (200)

(CD3OD) d(ppm): 7.24-7.11 (m, 9H), 3.35 (s, 2H), 2.60 (t, 4H, J=7.6 Hz),1.89 (m, 2H). LRMS: 269.1 (calc) 270.1 (found).

-   t) N-hydroxy-4-(5-phenylpentyl)benzamide (201)

(CD3OD) d(ppm) 1H, 7.64 (d, J=7.6 Hz, 2H), 7.22 (m, 4H), 7.12 (m, 3H),2.63 (t, J=7.6 Hz, 2H), 2.57 (t, J=7.2 Hz, 2H), 1.63 (m, 4H), 1.35 (m,2H). LRMS (ESI): (calc.) 283.4 (found) 284.3 (MH)+.

Example 45N-(1-aminocyclopropanecarbonyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamidehydrochloride (174a) Step 1:tert-butyl-1-1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)cyclopropylcarbamate

N-hydroxy-2-(4-(4-phenylbutyl)phenyl)acetamide 174 (1 equiv, 156 mg,0.55 mmol) was dissolved in DMF (3 mL).1-(tert-butoxycarbonylamino)cyclopropanecarboxylic acid (1.5 equiv, 166mg, 0.83 mmol) was then added followed by the sequential addition ofHOBt (1 equiv, 74 mg, 0.55 mmol) and EDC (1.5 equiv, 158 mg, 0.83 mmol).The reaction was then stirred overnight. The reaction was thenpartitioned between EtOAc (5 mL) and H₂O (5 mL). The organic phase wasseparated, dried over Na₂SO₄, filtered and concentrated. The residue waspurified by trituration with Et₂O to affordtert-butyl-1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)cyclopropylcarbamatein 166 mg (65%). LRMS: 466.5 (calc) 489.3 (MNa)⁺.

Step 2:N-(1-aminocyclopropanecarbonyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamidehydrochloride (174a)

A 4M solution of HCl in dioxane (3 mL) was added totert-butyl-1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)cyclopropylcarbamate(166 mg, 0.36 mmol). The reaction was stirred for 1 h. The solvent wasevaporated under reduced pressure. The compound was triturated with Et₂Oto affordN-(1-aminocyclopropanecarbonyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamidehydrochloride (174a) in 142 mg (99%). (dDMSO) δ (ppm) ¹H, 12.36 (bs,1H), 8.82 (bs, 3H), 7.26-7.08 (m, 9H), 3.43 (s, 2H), 2.55 (m, 4H),1.55-1.49 (m, 8H). LRMS: 366.1 (calc) 365.4 (MH)⁻.

Example 46

The following additional compounds were prepared by procedures analogousto those described in the Examples 40-43 and 45:

-   a)    (S)—N-(2-amino-3-phenylpropanoyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide    hydrochloride (174b)

(DMSO) δ (ppm) 1H, 12.39 (bs, 1H), 8.47 (bs, 3H), 7.31-6.96 (m, 1H),4.54 (m, 1H), 3.46 (s, 2H), 3.15 (m, 2H), 2.56 (q, J=6.8, 13.6 Hz, 4H),1.54 (m, 4H). LRMS (ESI): (calc.) 430.2 (found) 431.4 (MH)⁺.

-   b)    (S)—N-(2-amino-3-methylbutanoyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide    hydrochloride (174c)

(DMSO) δ (ppm) 1H, 12.32 (bs, 1H), 8.33 (bs, 3H), 7.24-7.09 (m, 9H),4.13 (m, 1H), 3.45 (s, 2H), 2.57 (m, 4H), 2.18 (m, 1H), 1.55 (m, 4H),1.03-0.94 (m, 6H). LRMS (ESI): (calc.) 382.2 (found) 383.1 (MH)⁺.

-   c)    (S)—N-(2-amino-3,3-dimethylbutanoyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide    hydrochloride (174d)

(dDMSO) δ (ppm) 1H, 12.49 (bs, 1H), 8.59 (bs, 3H), 7.26-7.09 (m, 9H),3.98 (s, 1H), 3.47 (s, 2H), 2.56 (m, 4H), 1.55 (m, 4H), 1.05 (s, 9H).LRMS (ESI): (calc.) 396.2 (found) 397.5 (MH)⁺.

-   d)    N-(1-aminocyclobutanecarbonyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide    hydrochloride (174e)

(dDMSO) δ (ppm) 1H, 12.52 (bs, 1H), 8.90 (bs, 1H), 7.19 (m, 9H), 3.47(s, 2H), 2.56 (m, 6H), 2.05 (m, 2H), 1.54 (m, 4H). LRMS (ESI): (calc.)380.2 (found) 381.4 (MH)⁺.

-   e)    N-(2-amino-2-methylpropanoyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide    hydrochloride (174f)

(dDMSO) δ (ppm) 1H, 12.45 (bs, 1H), 8.77 (bs, 3H), 7.16 (m, 9H), 3.45(s, 2H), 2.56 (m, 4H), 1.54 (m, 10H). LRMS (ESI): (calc.) 368.2 (found)369.4 (MH)⁺.

-   f)    N-(1-(aminomethyl)cyclopropanecarbonyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide    hydrochloride (174g)

(dDMSO) δ (ppm) 1H, 12.17 (bs, 1H), 8.00 (bs, 3H), 7.26-7.08 (m, 9H),3.42 (s, 2H), 3.05 (s, 2H), 2.55 (m, 4H), 1.54 (m, 4H), 1.35 (m, 2H),1.26 (m, 2H) LRMS (ESI): (calc.) 380.2 (found) 381.5 (MH)⁺

Example 47

The following additional prodrugs according to the present inventionwere also prepared.

-   (S)—N-(2,6-diaminohexanoyloxyl-2-(4-(4-phenylbutyl)phenyl)acetamide

LRMS (ESI): (calc.) 411.25 (found) 412.509 (MH)+

(DMSO) d(ppm) 1H, 12.50 (s, 1H), 8.66 (s, 3H), 7.81 (s, 3H), 7.27-7.10(m, 9H), 4.27-4.21 (m, 1H), 3.48-3.46 (m, 2H), 2.75-2.68 (m, 2H),2.60-2.52 (m, 4H), 1.88-1.80 (m, 2H), 1.60-1.40 (m, 8H).

-   N-(2-hydroxyacetoxy)-2-(4-(4-phenylbutyl)phenyl)acetamide

LRMS (ES−): (calc.) 341.16 (found) 340.466 (MH)+

(DMSO) d(ppm) 1H, 11.95 (s, 1H), 7.27-7.09 (m, 9H), 5.64 (t, J=6.4 Hz,1H), 4.17 (d, J=6.4 Hz, 2H), 3.42 (s, 2H), 2.60-2.52 (m, 4H), 1.58-1.53(m, 4H)

-   (S)—N-(2-amino-5-guanidinopentanoyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide

LRMS (ESI): (calc.) 439.26 (found) 440.651 (MH)+

(DMSO) d(ppm) 1H, 12.60 (s, 1H), 8.77 (s, 1H), 8.43 (s, 4H), 8.00 (s,1H), 7.85 (s, 1H), 7.37-7.12 (m, 9H), 3.95-3.85 (m, 1H), 3.20-3.10 (m,4H), 2.60-2.52 (m, 2H), 1.92-1.72 (m, 4H), 1.70-1.45 (m, 4H)

-   (S)-2,6-diamino-N-(1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)cyclopropyl)hexanamide

LRMS (ESI): (calc.) 494.29 (found) 495.592 (MH)+

(DMSO) d(ppm) 1H, 12.08 (s, 1H), 9.36 (s, 1H), 8.23 (s, 3H), 7.86 (s,3H), 7.27-7.08 (m, 9H), 3.75-3.65 (m, 1H), 3.39 (s, 2H), 2.78-2.67 (m,2H), 2.62-2.52 (m, 4H), 1.80-1.66 (m, 2H), 1.64-1.46 (m, 8H), 1.44-1.28(m, 2H), 1.38-1.26 (m, 2H)

-   N-(1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)cyclopropyl)nicotinamide

LRMS (ESI): (calc.) 471.22 (found) 472.525 (MH)+

(DMSO) d(ppm) 1H, 9.45 (s, 1H), 9.00 (s, 1H), 8.72 (d, 1H, J=4.8 Hz),8.19 (d, 1H, J=8 Hz), 7.51 (dd, 1H, J=8 Hz, 4.8 Hz), 7.27-7.05 (m, 9H),2.59-2.53 (m, 4H), 1.60-1.50 (m, 4H), 1.32-1.27 (m, 2H), 1.25-1.21 (m,2H)

-   (S)-2-amino-3-methyl-N-(1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)cyclopropyl)butanamide

LRMS (ESI): (calc.) 465.26 (found) 466.695 (MH)+

(DMSO) d(ppm) 1H, 12.04 (s, 1H), 9.21 (s, 1H), 8.09 (s, 3H), 7.27-7.06(m, 9H), 3.50-3.45 (m, 1H), 3.40-3.37 (m, 2H), 2.61-2.53 (m, 4H),2.14-2.04 (m, 1H), 1.62-1.48 (m, 6H), 1.24-1.16 (m, 2H), 0.98-0.90 (m,6H)

-   (S)-2-amino-3-phenyl-N-(1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)cyclopropyl)propanamide

LRMS (ESI): (calc.) 513.26 (found) 514.675 (MH)+

(DMSO) d(ppm) 1H, 12.03 (s, 1H), 9.19 (s, 1H), 8.14 (s, 3H), 7.29-7.08(m, 14H), 3.92-3.85 (m, 1H), 3.40-3.37 (m, 2H), 3.06-2.94 (m, 2H),2.60-2.53 (m, 4H), 1.57-1.51 (m, 4H), 1.51-1.46 (m, 2H), 1.40-0.80 (m,2H)

-   2-(4-(4-phenylbutyl)phenyl)-N-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yloxy)acetamide

LRMS (ESI): (calc.) 445.2 (found) 446.6 (MH)+

(CD3OD) d(ppm) 1H, 7.24-7.10 (m, 9H), 4.50 (d, J=8.0 Hz, 1H), 3.90 (dd,J=2.4, 12.0 Hz, 1H), 3.65 (dd, J=6.0, 11.6 Hz, 1H), 3.40 (s, 2H),3.35-3.25 (m, 3H), 2.61 (m, 4H), 1.62 (m, 4H)

-   N-(2,3-dihydroxypropanoyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide

LRMS (ESI): (calc.) 371.17 (found) 000.0 (M)-370.478

(CD3OD) d(ppm) 1H, 7.25-7.08 (m, 9H), 4.41 (t, J=4 HZ, 1H), 3.81 (d, J=4Hz, 2H), 3.51 (s, 2H), 2.64-2.58 (m, 4H), 1.65-1.58 (m, 4H)

-   N-hydroxy-2-(4-(4-(3-morpholinophenyl)butyl)phenyl)acetamide

LRMS (ESI): (calc.) 368.21 (found) 369.571 (MH)+

(dmso) d(ppm) 1H, 10.62 (s, 1H), 8.78 (s, 1H), 7.14-7.04 (m, 5H),6.72-6.68 (m, 2H), 6.60 (d, J=7.2 Hz, 1H), 3.70 (t, J=4.8 Hz, 4H), 3.20(s, 2H), 3.04 (t, J=4.8 Hz, 4H), 2.56-2.49 (m, 4H), 1.58-1.48 (m, 4H)

-   N-hydroxy-2-phenyl-2-(4-(4-phenylbutyl)phenyl)acetamide

LRMS (ESI): (calc.) 359.2 (found) 360.4 (MH)+

(CD3OD) d(ppm) 1H, 7.30-7.11 (m, 14H), 4.73 (s, 1H), 2.60 (m, 4H), 1.61(m, 4H)

LRMS (ESI): (calc.) 431.5 (found) 431.5 (M)+

(CD3OD) d(ppm) 1H, 8.84 (s, 1H), 8.71 (d, J=6.0 Hz, 1H), 8.49 (d, J=8.0Hz, 1H), 7.96 (t, J=6.8 Hz, 1H), 7.24-7.10 (m, 9H), 4.30 (s, 3H), 3.47(s, 2H), 3.22 (t, J=7.2 Hz, 2H), 2.97 (t, J=6.8 Hz, 2H), 2.61 (m, 4H),1.62 (m, 4H)

-   2-(4-(4-(fluorophenyl)butyl)phenyl)-N-hydroxyamide

LRMS (ESI): (calc.) 301.1 (found) 302.3 (MH)+

(dDMSO) d(ppm) 1H, 7.19 (m, 2H), 7.11-6.98 (m, 6H), 3.02 (s, 2H), 2.54(m, 4H), 1.53 (m, 4H)

LRMS (ESI): (calc.) 421.2 (found) 421.5 (MH)+

(CD3OD) d(ppm) 1H, 12.13 (bs, 1H), 10.11 (s, 1H), 9.14 (s, 1H),7.27-7.09 (m, 9H), 4.67 (t, J=6.0 Hz, 2H), 3.86 (s, 3H), 3.39 (s, 2H),3.17 (t, J=6.0 Hz, 2H), 2.56 (m, 4H), 1.55 (m, 4H)

LRMS (ESI): (calc.) 403.2 (found) 403.4 (M)+.

(CD3OD) d(ppm) 1H, 9.66 (s, 1), 9.14 (m, 2), 8.25 (m, 1H), 7.26-7.10 (m,9H), 4.49 (s, 3H), 3.60 (s, 2H), 2.62 (m, 4H), 1.64 (m, 4H)

Example 48 Inhibition of Histone Deacetylase Enzymatic Activity

HDAC inhibitors are screened against histone deacetylase enzyme innuclear extracts prepared from the human small cell lung cancer cellline H446 (ATTC HTB-171) and against a cloned recombinant human (e.g.,HDAC-1) enzyme expressed and purified from a Baculovirus insect cellexpression system.

For deacetylase assays, 20,000 cpm of the [³H]-metabolically labeledacetylated histone substrate (M. Yoshida et al., J. Biol. Chem. 265(28):17174-17179 (1990)) is incubated with 30 mg of H446 nuclear extract oran equivalent amount of the cloned recombinant hHDAC-1 for 10 minutes at37° C. The reaction is stopped by adding acetic acid (0.04 M, finalconcentration) and HCl (250 mM, final concentration). The mixture isextracted with ethyl acetate and the released [³H]-acetic acid wasquantified by scintillation counting. For inhibition studies, the enzymeis preincubated with compounds at 4° C. for 30 minutes prior toinitiation of the enzymatic assay. IC₅₀ values for HDAC enzymeinhibitors are determined by performing dose response curves withindividual compounds and determining the concentration of inhibitorproducing fifty percent of the maximal inhibition.

Alternatively (Table 4a), the following protocol is used to assay thecompounds of the invention. In the assay, the buffer used is 25 mMHEPES, pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl₂ and the substrate isBoc-Lys(Ac)-AMC in a 50 mM stock solution in DMSO. The enzyme stocksolution is 4.08 μg/mL in buffer. The compounds are pre-incubated (2 μlin DMSO diluted to 13 μl in buffer for transfer to assay plate) withenzyme (20 μl of 4.08 μg/ml) for 10 minutes at room temperature (35 μlpre-incubation volume). The mixture is pre-incubated for 5 minutes atroom temperature. The reaction is started by bringing the temperature to37° C. and adding 16 μl substrate. Total reaction volume is 50 μl. Thereaction is stopped after 20 minutes by addition of 50 μl developer,prepared as directed by Biomol (Fluor-de-Lys developer, Cat. #KI-105). Aplate is incubated in the dark for 10 minutes at room temperature beforereading (λ_(Ex)=360 nm, λ_(Em)=470 nm, Cutoff filter at 435 nm).

Representative data are presented in Tables 4 and 4a. In the firstcolumn of Table 4 are reported IC₅₀ values determined against histonedeacetylase in nuclear extracts from H446 cells (pooled HDACs). In thesecond column of Table 4 are reported IC₅₀ values determined againstrecombinant human HDAC-1 enzyme (rHDAC-1). For less active compounds,the data are expressed as the percent inhibition at the specifiedconcentration.

TABLE 4 Inhibition of Histone Deacetylase pooled HDACs rHDAC-1 ExampleCpd. Structure IC₅₀ (μM) IC₅₀ (μM) Ex. 1  4

7 Ex. 2  7

70 Ex. 3  8

15 Ex. 4  9

9 Ex. 5  10

30 Ex. 6  11

10 Ex. 7  12

3 Ex. 8  13

0.9 Ex. 9  14

36% @ 100 μM Ex. 10  15

25 Ex. 11  16

38% 100 μM Ex. 12  17

47% 100 μM Ex. 13  18

160 Ex. 14  19

20 Ex. 15  26

<20 Ex. 16  32

<20 Ex. 17  34

2 0.3 Ex. 18  36

0.5 0.2 Ex. 19  38

0.75 0.1 Ex. 20  42

5 1.0 Ex. 21  45

4 Ex. 22  50

5 Ex. 23  53

25 Ex. 24  56

15 Ex. 25  61

4 Ex. 26  64

12% @ 100 μM Ex. 27  68

 3% @ 20 μM Ex. 28  70

5.5 0.9 Ex. 28  71

44% @ 20 μM Ex. 28  73

35% @ 20 μM Ex. 29  77

□ 0.65 Ex. 30  81

>50 >25 Ex. 31  86

3.8 Ex. 31  87

3 0.6 Ex. 31  88

0.6 0.075 Ex. 31  89

3 0.9 Ex. 31  90

0.4 0.09 Ex. 31  91

5 2 Ex. 31  92

>20 17 Ex. 31  93

0.35 0.05 Ex. 31  94

0.4 0.03 Ex. 31  95

0.8 0.2 Ex. 31  96

33% @ 5 μM Ex. 31  97

0.8 0.28 Ex. 31  98

0.55 0.06 Ex. 31  99

0.9 0.05 Ex. 31 100

0.8 0.75 Ex. 31 101

0.3 0.04 Ex. 31 102

5.5 0.8 Ex. 31 103

0.7 0.05 Ex. 31 104

21% @ 5 μM Ex. 31 105

0.55 0.2 Ex. 31 106

0.8 0.3 Ex. 31 107

 0% @ 1 μM 5 Ex. 31 108

10% @ 1 μM 0.3 Ex. 31 109

32% @ 1 μM 0.12 Ex. 31 110

0.7 0.55 Ex. 31 111

0.4 0.095 Ex. 31 112

1.2 0.6 Ex. 31 113

46% @ 1 μM 0.2 Ex. 31 114

40% @ 1 μM 0.1 Ex. 31 115

53% @ 1 μM 0.1 Ex. 31 116

4 Ex. 31 117

 0% @ 20 μM 1.9 Ex. 31 118

 0% @ 20 μM 2.3 Ex. 31 119

3 Ex. 31 120

0.12 0.01 Ex. 31 121

23 Ex. 31 122

2.3 Ex. 31 123

1 Ex. 32 128

0.3 Ex. 32 129

3.0 Ex. 33 136

9 0.5 Ex. 34 139

44% @ 20 μM Ex. 34 143

55% @ 20 μM 2.4 Ex. 34 144

 6% @ 20 μM 6.9 Ex. 35 145

3.8 0.84 Ex. 35 146

2.9 0.91 Ex. 35 147

1.9 0.48 Ex. 36 148

5 2.0 Ex. 36 149

 8% @ 20 μM 0.1 Ex. 36 150

10 1.0 Ex. 36 151

7.5 2.3 Ex. 36 152

35% @ 20 μM Ex. 36 153

5 4.8 Ex. 36 154

□ 0.9 Ex. 36 155

39% @ 20 μM Ex. 36 156

5 0.75 Ex. 36 157

6 2.4 Ex. 36 158

>20 Ex. 36 159

1.5 Ex. 36 160

1.2 Ex. 36 161

0.05 Ex. 36 162

0.04 Ex. 37 164

5.0 Ex. 37 165

2.0 Ex. 37 166

Ex. 37 167

Ex. 38 168

 0% @ 20 μM 3 Ex. 39 170

48% @ 2 μM 0.57 171

20 172

10 173

35% @ 20 μM 174

>20 175

>2 176

20% @ 20 μM 177

10% @ 20 μM 178

 2% @ 20 μM >20

TABLE 4a rHDAC-1 rHDAC-8 Cpd. Structure IC₅₀ (μM) IC₅₀ (μM) 182

0.7 PI (64%) 1.3 PI (74%) 179

0.5  0.53 183

0.38 0.41 184

0.93 0.4 PI (79%) 185

0.84 0.61 187

0.2 PI (62%) 0.21 PI = partial inhibition (100% inhibition not reached −PI value indicates IC₅₀ (at maximum % inhibition reached))

Example 49 Inhibition of Histone Deacetylase in Whole Cells 1. HistoneH4 Acetylation in Whole Cells by Immunoblots

T24 human bladder cancer cells growing in culture are incubated withHDAC inhibitors for 16 hours. Histones are extracted from the cellsafter the culture period as described by M. Yoshida et al. (J. Biol.Chem. 265(28): 17174-17179 (1990)). 20 μμg of total histone protein areloaded onto SDS/PAGE and transferred to nitrocellulose membranes.Membranes are probed with polyclonal antibodies specific for acetylatedhistone H-4 (Upstate Biotech Inc.), followed by horse radish peroxidaseconjugated secondary antibodies (Sigma). Enhanced Chemiluminescence(ECL) (Amersham) detection is performed using Kodak films (EastmanKodak). Acetylated H-4 signal is quantified by densitometry.

Data for selected compounds are presented in Table 5. Data are presentedas the concentration effective for reducing the acetylated H-4 signal by50% (EC₅₀).

TABLE 5 Inhibibition of Histone Acetylation in Cells Cpd. Structure EC₅₀(μM)  36

5  90

1  98

1 107

5 118

3 120

1 122

2

2. Acid Urea Triton (AUT) Gel Analysis of Histone Acetylation.

Human cancer cells (T24, 293T or Jurkat cells) growing in culture areincubated with HDAC inhibitors for 24 h. Histones are extracted from thecells as described by M. Yoshida et al. (J. Biol. Chem. 265(28):17174-17179 (1990)). Acid urea triton (AUT) gel electrophoresis is usedfor detection of acetylated histone molecules. Histones (150 μμg oftotal protein) are electrophoresed at 80 V for 16 h at room temperatureas described by M. Yoshida et al., supra. Gels are stained withCoomassie brilliant blue to visualize histones, dried and scanned bydensitometry to quantified acetylation of histones.

Example 50 Antineoplastic Effect of Histone Deacetylase Inhibitors onTumor Cells In Vivo

Eight to ten week old female BALB/c nude mice (Taconic Labs, GreatBarrington, N.Y.) are injected subcutaneously in the flank area with2×10⁶ preconditioned A549 human lung carcinoma cells. Preconditioning ofthese cells is done by a minimum of three consecutive tumortransplantations in the same strain of nude mice. Subsequently, tumorfragments of approximately 30 mgs are excised and implantedsubcutaneously in mice, in the left flank area, under Forene anesthesia(Abbott Labs, Geneve, Switzerland). When the tumors reach a mean volumeof 100 mm³, the mice are treated intravenously, subcutaneously, orintraperitoneally by daily injection, with a solution of the histonedeacetylase inhibitor in an appropriate vehicle, such as PBS,DMSO/water, or Tween 80/water, at a starting dose of 10 mg/kg. Theoptimal dose of the HDAC inhibitor is established by dose responseexperiments according to standard protocols. Tumor volume is calculatedevery second day post infusion according to standard methods (e.g.,Meyer et al., Int. J. Cancer 43: 851-856 (1989)). Treatment with theHDAC inhibitors according to the invention causes a significantreduction in tumor weight and volume relative to controls treated withsaline only (i.e., no HDAC inhibitor). In addition, the activity ofhistone deacetylase when measured is expected to be significantlyreduced relative to saline treated controls.

Example 51 Synergistic Antineoplastic Effect of Histone DeacetylaseInhibitors and Histone Deacetylase Antisense Oligonucleotides on TumorCells In Vivo

The purpose of this example is to illustrate the ability of the histonedeacetylase inhibitor of the invention and a histone deacetylaseantisense oligonucleotide to synergistically inhibit tumor growth in amammal. Preferably, the antisense oligonucleotide and the HDAC inhibitorinhibit the expression and activity of the same histone deacetylase.

As described in Example 10, mice bearing implanted A549 tumors (meanvolume 100 mm³) are treated daily with saline preparations containingfrom about 0.1 mg to about 30 mg per kg body weight of histonedeacetylase antisense oligonucleotide. A second group of mice is treateddaily with pharmaceutically acceptable preparations containing fromabout 0.01 mg to about 5 mg per kg body weight of HDAC inhibitor.

Some mice receive both the antisense oligonucleotide and the HDACinhibitor. Of these mice, one group may receive the antisenseoligonucleotide and the HDAC inhibitor simultaneously intravenously viathe tail vein. Another group may receive the antisense oligonucleotidevia the tail vein, and the HDAC inhibitor subcutaneously. Yet anothergroup may receive both the antisense oligonucleotide and the HDACinhibitor subcutaneously. Control groups of mice are similarlyestablished which receive no treatment (e.g., saline only), a mismatchantisense oligonucleotide only, a control compound that does not inhibithistone deacetylase activity, and a mismatch antisense oligonucleotidewith a control compound.

Tumor volume is measured with calipers. Treatment with the antisenseoligonucleotide plus the histone deacetylase protein inhibitor accordingto the invention causes a significant reduction in tumor weight andvolume relative to controls.

Example 52 Solubility

The solubility of several compounds according to the invention wasmeasured, and the results are displayed in the table below.

Solubility (μM) Q pH 2 Water* H 3 3

4 5

132 195

10 9

>250 >250

142 136

186 130

49

>2000 >1000

166 165 *Milli-Q  ® purified water

While the invention has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations of the invention following, in general, theprinciples of the invention and including such departures from thepresent disclosure as come within known or customary practice within theart to which the invention pertains and as may be applied to theessential features hereinbefore set forth, and as follows in the scopeof the appended claims.

1.-34. (canceled)
 35. A compound selected from the group consisting ofN-hydroxy-2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetamide;2-(4-(4-(benzo[c][1,2,5]oxadiazol-5-yl)but-3-ynyl)phenyl)-N-hydroxyacetamide;N-Hydroxy-2-(3-(4-phenylbutyl)phenyl)acetamide;N-hydroxy-2-(4-(4-(4-(trifluoromethyl)phenyl)butyl)phenyl)acetamide;2-(4-(4-(1H-indol-5-yl)butyl)phenyl)-N-hydroxyacetamide;N-hydroxy-2-(4-(4-(3-(trifluoromethyl)phenyl)butyl)phenyl)acetamide;N-hydroxy-2-(4-(4-(2-(trifluoromethyl)phenyl)butyl)phenyl)acetamide;N-hydroxy-2-(4-(4-(imidazo[1,2-a]pyridin-6-yl)butyl)phenyl)acetamide;2-(4-(4-(benzo[d][1,3]dioxol-5-yl)butyl)phenyl)-N-hydroxyacetamide;N-hydroxy-2-(4-(5-phenylpentyl)phenyl)acetamide;N-hydroxy-2-(4-(4-(pyridin-4-yl)butyl)phenyl)acetamide;N-hydroxy-2-(4-(4-(pyridin-3-yl)butyl)phenyl)acetamide;N-hydroxy-2-(4-(4-(pyridin-2-yl)butyl)phenyl)acetamide;N-hydroxy-2-(4-(3-phenylpropyl)phenyl)acetamide; andN-hydroxy-4-(5-phenylpentyl)benzamide; and pharmaceutically acceptablesalts thereof.
 36. A compound selected from the group consisting of

and pharmaceutically acceptable salts thereof.
 37. A compositioncomprising a compound of claim 35 or a pharmaceutically acceptable saltthereof.
 38. A composition comprising a compound of claim 36 or apharmaceutically acceptable salt thereof.